Is Fasciola monospecies? : Cross experiments between Fasciola hepatica and F.gigantica

• Project/Area Number: 14560243
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Basic veterinary science/Basic zootechnical science
• Research Institution: Iwate University
• Principal Investigator: ITAGAKI Tadashi Iwate University, Veterinary Parasitology, Associate Professor, 農学部, 助教授 (80203074)
• Project Period (FY): 2002 – 2004
• Project Status: Completed (Fiscal Year 2004)
• Budget Amount: ¥2,800,000 (Direct Cost: ¥2,800,000); Fiscal Year 2004: ¥700,000 (Direct Cost: ¥700,000); Fiscal Year 2003: ¥700,000 (Direct Cost: ¥700,000); Fiscal Year 2002: ¥1,400,000 (Direct Cost: ¥1,400,000)
• Keywords: liver fluke / Fasciola hepatica / Fasciola gigantica / parthenogenic diploid / parthenogenic triploid / cross experiments / DNA / ITS / 感染実験

Research Abstract

The first purpose of the study is to find small experimental animals susceptible to Fasciola hepatica and F.gigantica through experimental infections. Rats, gerbils (Meriones unguiculatus), and the rat-like hamster, Tscherskia triton were orally inoculated with 2 to 10 metacercariae of the two species, and then sacrificed after 2 to 3 months. The animals were checked for the presence of flukes in the liver parenchyma and bile duct, infection rate and damage of the liver. The flukes obtained were fixed in 70% ethanol, stained with Carmin solution, and measured on the body length and width. F.hepatica had high recovery rate and matured in the three animal species, while F.gigantica had low rate and did not matured in all the hosts. These findings showed that these three animals could not play the role of suitable hosts of both Fasciola species.
Cross experiments between bisexual F.hepatica (Fh) and parthenogenic diploid (2n) or triploid (3n) were carried out using rodent hosts, and the fi rst filial generations (F1) were genetically characterized by analyzes of chromosomes and DNA sequences. In the first experiments, single metacercariae of both Fh and 2n were orally inoculated to the rat-like hamster, Tscherskia triton. A 2n fluke recovered from the host was confirmed to conjugate with a Fh fluke by morphological observation. F1 produced from the 2n fluke was allopolyploid (triploid) and this suggested that Fh and 2n were different species each other. In the second experiments, 10 metacercariae each of Fh and 3n were inoculated to Wistar rats. The 3n flukes recovered from the rats and confirmed to conjugate with a Fh fluke were provided for production of F1. Genetic observation showed that the F1 was a tetraploid and had 10 univalent chromosomes and 10 trivalent chromosomes in reproductive cells. Furthermore, the F1 had abnormal spermatogenesis. These findings suggested that F.hepatica and parhtenogenic triploid were different species each other. The F1 tetraploid was sterile.
Cross experiments between Fh and Fg, and Fg and parthenogenic diploid (2n) or triploid (3n) were carried out using goat hosts. Unfortunately, the goat infected with both Fh and Fg was dead by the time the flukes have matured and therefore the F1 flukes between the two species could not be obtained. While the F1s between Fg and the parthenogenic plolpoids have not been obtained by the time the experimental period expired. However, these findings suggest that F.hepatica, F.gigantica and parthenogenic Fasciola sp. are different species each other.

Research Products

• [Journal Article] Prevalence of Giardia intestinalis infection in dogs of breeding Kennels in Japan. (2005)
  • Author(s): Itoh, N., N.Muraoka, H.Saeki, M.Aoki, T.Itagaki
  • Journal Title: J.Vet.Med.Sci. 印刷中
  • NAID: 110003963797
• [Journal Article] 日本産肝蛭の遺伝的特徴および起源について-リボソームDNAおよびミトコンドリアDNAからみた一考察- (2004)
  • Author(s): 板垣 匡, 紀川将之, 坂口圭介, 寺崎邦生, 柴原壽行, 福川孝
  • Journal Title: 獣医寄生虫誌 3 Pages: 29-35
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Genetic characterization and origin of Japanese Fasciola sp. based on the sequences of ribosomal and mitochondrial DNA. (2004)
  • Author(s): Itagaki, T., Kikawa, M., Sakaguchi, K., Terasaki, K., Shibahara, T., Fukuda, K.
  • Journal Title: Jpn.J.Vet.Parasitol. 3 Pages: 29-35
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] 日本産肝蛭の遺伝的特徴および起源について-リボソームDNAおよびミトコンドリアDNAからみた一考察- (2004)
  • Author(s): 板垣 匡, 紀川将之, 坂口圭介, 寺崎邦生, 柴原壽行, 福田孝一
  • Journal Title: 獣医寄生虫誌 3 Pages: 29-35
• [Journal Article] Treatment of Notoedres cati infestation in cats with selamectin. (2004)
  • Author(s): Itoh, N., N.Muraoka, M.Aoki, T.Itagaki
  • Journal Title: Veterinary Record 154 Pages: 409-409
• [Journal Article] Prevalence of Toxocara canis infection in household dogs. (2004)
  • Author(s): Itoh, N., N.Muraoka, M.Aoki, T.Itagaki
  • Journal Title: J.Jpn.Assoc.Inf.Dis. 78 Pages: 114-119
  • NAID: 10012958683
• [Journal Article] 子犬におけるELISAによるジアルジア抗原の検出状況 (2004)
  • Author(s): 伊藤直之, 村岡 登, 青木美樹子, 板垣 匡
  • Journal Title: 日獣会誌 57 Pages: 579-582
  • NAID: 10013518215
• [Journal Article] Experimental fascioliasis in the rat-like hamster, Tscherskia triton, and other rodent hosts. (2003)
  • Author(s): Terasaki, K., Noda, Y., Shibahara, T., Itsaki.T., Fukuda, K., Tsuchiya, K.
  • Journal Title: Parasitol.Int. 52 Pages: 147-154
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] ウルグアイ産Fasciola hepaticaの配偶子形成と受精 (2003)
  • Author(s): 寺崎邦生, 柴原壽行, 板垣 匡, 福田孝, 野川安孝, 峯みゆき, 本村亜希, 由本真由佳
  • Journal Title: 獣医畜産新報 56 Pages: 97-103
  • NAID: 50000743276
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Experimental fascioliasis in the rat-like hamster, Tscherskia triton, and other rodent hosts. (2003)
  • Author(s): Terasaki, K., Noda, Y., Shibahara, T., Itagaki, T., Fukuda, K, Tsuchiya, K.
  • Journal Title: Parasitol.Int. 52 Pages: 147-154
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Gametogenesis and fertilization of Fasciola hepatica from Uruguay. (2003)
  • Author(s): Terasaki, K., Shibahara, T., Itagaki, T., Fukuda, K., Noda, Y., Mine, M., Motomura, A., Yamamoto, M.
  • Journal Title: J.Vet.Med.(Tokyo) 56 Pages: 97-103
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The rat-like hamster, Tscherskia triton, as a host for the parthenogenetic liver fluke Fasciola sp. (2002)
  • Author(s): Terasaki, K., Noda, Y., Itagaki, T., Shibahara, T., Tsuchiya, K., Fukuda, K., Inoue, Y., Iwainura, M., Hino, A.
  • Journal Title: J.Parasitol. 88 Pages: 1035-1037
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The rat-like hamster, Tscherskia triton, as a host for the parthenogenetic liver fluke Fasciola sp. (2002)
  • Author(s): Terasaki, K., Noda, Y., Itagaki, T., Shibahara, T., Tsuchiya, K., Fukuda K., Inoue, Y., Iwamura, M, Hino, A.
  • Journal Title: J.Parasitol. 88 Pages: 1035-1037
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Genetic characterization of parthenogenic Fasciola sp.in Japan on the basis of the sequences of ribosonial and mitochondrial DNA.
  • Author(s): Itagaki, T., Kikawa, M., Sakaguchi, K., Shimo, J., Terasaki, K., Shibahara, T, Fukuda, K.
  • Journal Title: Parasitology (In press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Molecular Cacharacterization of parthenogenic Fasciola sp.in Korea on the basis of DNA sequences of ribosomal ITSland mitochondrial NDI gene.
  • Author(s): Itagaki, T, Terasaki, K., ShibKikawa, M., Shibahara, T., Fukucda, K.
  • Journal Title: J.Vet.Med.Sci. (In press)
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Genetic characterization of parthenogenic Fasciola sp. in Japan on the basis of the sequences of ribosomal and mitochondrial DNA.
  • Author(s): Itagaki, T., Kikawa, M., Sakaguchi, K., Shimo, J., Terasaki, K., Shibahara, T., Fukuda, K.
  • Journal Title: Parasitology (in press)
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Molecular characterization of parthenogenic Fasciola sp. in Korea on the basis of DNA sequences of ribosomal ITS1 and mitochondrial NDI gene.
  • Author(s): Itagaki, T., Kikawa, M., Terasaki, K., Shibahara, T., Fukuda, K.
  • Journal Title: J.Vet.Med.Sci. (in press)
  • NAID: 110004049152
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Terasaki, K., Noda, Y., Itagaki, T., et al.: "The rat like hamster, Tscherskia triton as host for the parthenogenetic liver fluke Fasciola sp."J.Parasitiol.. 88. 1035-1037 (2002)
• [Publications] Terasaki, K., Noda, Y., Shibahara, Y., Itagaki, T., Fukuda, K., Tsuchiya, K.: "Experimental fascioliasis in the rat-like hamster, Tscherskia triton, and other rodent hosts"Paral.Int.. 52. 147-154 (2003)
• [Publications] 寺崎邦生, 柴原壽行, 板垣 匡: "ウルグアイ産Fasciola hepaticaの配偶子形成と受精"獣医畜産新報. 56. 97-103 (2003)
• [Publications] Itagaki, T., Tsumagari, N., Tsutsumi, K., Chinone, S.: "Discrimination of three amphistome species by PCR-RFLP based on rDNA ITS2 markers"J.Vet.Med.Sci.. 65. 931-933 (2003)
• [Publications] Ito, M., Itagaki, T.: "Survey on wild rodents for endoparasites in Iwate Prefecture, Japan"J.Med.Sci.. 65. 1151-1153 (2003)