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Clinical pathobiological studies on Sarcocystosis

Research Project

Project/Area Number 04454120
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Applied veterinary science
Research InstitutionThe University of Tokyo

Principal Investigator

ONO Kenichiro  Fac.of Agr., Univ.of Tokyo Prof., 農学部, 教授 (50111480)

Co-Investigator(Kenkyū-buntansha) MATSUKI Naoaki  Fac.of Agr., Univ.of Tokyo Ass.Prof., 農学部, 助手 (40251417)
NAKAYAMA Hiroyuki  Fac.of Agr., Univ.of Tokyo Aso.Prof., 農学部, 助教授 (40155891)
KONO Michiko  Fac.of Agr., Univ.of Tokyo Ass.Prof., 農学部, 助手 (70092202)
HASEGAWA Atsuhiko  Fac.of Agr., Univ.of Tokyo Prof., 農学部, 教授 (90011923)
SUZUKI Naoyoshi  Proto.Res.Center, Obihiro Univ.Prof., 畜産学部, 教授 (10003071)
後飯塚 僚  東京大学, 農学部, 助手 (60205581)
Project Period (FY) 1992 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥6,600,000 (Direct Cost: ¥6,600,000)
Fiscal Year 1993: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1992: ¥5,200,000 (Direct Cost: ¥5,200,000)
KeywordsSarcocystis / Sporozoites / Purification / Enzyme activity / Isozyme / Membrane protein / Excystation / 抗原特異性 / 形態学的変化 / ザルコンステス
Research Abstract

Clinical pathological examination was performed in 4 Sarcocystis species(Sarcocystis arieticanis, S.capracanis, S.hircicanis, and S.ovicanis) by the enzymatic analysis.
1. In vitro excystation and cryoreservation of Sarcocystis species : A standard pretreatment was performed by preincubation of sporocysts in sodium hypochlorite(NaOCl) solution at room temperature. Then, sporocysts were incubated for 1 hr at 39 C in excystaion fluid(RPMI 1640, 10% FCS and 15% bovine bile). Additional sonication of pretreated sporocysts increased excystation rates. Excystaion rates for 1-9 months old sporocysts were 77% for S.capracanis, 77 % for S.hircicanis, 72 % for S.arieticanis and 92 % for S.ovicanis. Sporozoites were also subjected to cryopreservation in RPMI 1640 containing 10% FCS and 7.5% DMSO.After storage in liquid nitrogen, sporozoites proved to be infective for their intermediate hosts.
2. Purification of in vitro excysted Sarcocystis sporozoites : Excysted Sarcocystis sporozoites were purifi … More ed by DE 52 anion-exchange cellulose column. After passage of excysted sporozoites through a column, the mean recovery was 88% for S.capracanis, 71% for S.hircicanis, 77% for S.ovicanis, and 74% for S.arieticanis. Most of the sporozoites were mortile, and the parasite suspension was free of excystation debris.
3. Isoenzyme profiles of 4 Sarcocystis sporozoites : The 4 Sarcocystis sporozoites obtained by an optimized excystation techniques were lysed by freezing and thawing, followed by ultrasonocation. On enzyme screening assay for 16 enzymes related to carbohydrate metabolism. Lactae dehydrogenase(LDH), malate dehydrogenase(MDH), and glucose-6-phosphate dehydrogenase(G6PD) activities were detected in all 4 samples. On isoenzyme patterns of these 3 enzymes, all 4 Sarcocystis species showed 1 single band of LDH activity with a more anodical migration in S.ovicanis and S.capracaniis than in the remaining 2 species. In MDH activity, 1 single band was shown in S.hircicanis whereas a more diffuse band was detected at same position in S.arieticanis and multiple band was shown in S.ovicanis and S.capracanis. In G6PD activity, 2 narrow bands were detected in all 4 samples, however, no difference was observed on their migration distances. Less

Report

(3 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • Research Products

    (11 results)

All Other

All Publications (11 results)

  • [Publications] ONO,K.et al.: "Purification of in vitro excysted Sarcocystis sporozoites by passage through a modified DE-52 anion-exchange colunn." Parasitol.Res.77. 717-719 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] HORN,R.et al.: "In vitro excystation and cryoreservation of ovine and caprine Sarcocystis species." J.Protozoal.Res.1. 13-21 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] GREINER,M.et al.: "Comparative investigation of sporozoites of 4 Sarcocystis species by isoenzyme electrophoresis." J.Protozoal.Res.2. 123-127 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Ono. K.et al: "Purification of invitro excysted Sarcocystic sporozoites by passage through a modified DE-52 anion-exchange column." Parasitol.Res.77. 717-719 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Horn.K.et al: "In vitro excystation and cryoreservation of ovine and caprine Sarcocystis species." J.Protozool.Res.1. 13-21 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Greiner, M.et al: "Comparative investigation of sporozoites of 4 Sarcocystis species by isoenzyme electrophoresis." J.Protozool.Res.2. 123-127 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] ONO,K.et al.: "Purification of in vitro excysted Sarcocystis sporozoites by passage through a modified DE-52 anion-exchange column" Porasitol.Res.77. 717-719 (1991)

    • Related Report
      1993 Annual Research Report
  • [Publications] Horn,K.et.al.: "In vitno excystation and cryoreservution of ovine and caprine Sarcocystis species" J.Protozool.Res.1. 13-21 (1991)

    • Related Report
      1993 Annual Research Report
  • [Publications] Greiner,M.et al.: "Comparative in vestigation of sporozoites of 4 Sarcocystis species by issenzyme electrophoresis" J.Protozool.Res.2. 123-127 (1992)

    • Related Report
      1993 Annual Research Report
  • [Publications] Ono,K.et al.: "Purification of in vitro excysted Sarcocystis sporozoites by passage through a modififed DE 52 anion-exchange column" Parasitol.Res. 77. 717-719 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Horn,K.et al.: "In vitro excystation and cryoreservation of ovine and caprine Sarcoccystis species." J.Protozool.Res.1. 13-21 (1991)

    • Related Report
      1992 Annual Research Report

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Published: 1992-04-01   Modified: 2016-04-21  

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