Still and Moving Image Evidences for Mating of Echinococcus granulosus Reared in Culture Media
Abstract
Background: Echinococcus granulosus cultivation is very important for improvement of different aspect of medical and veterinary researches. Despite many advances in this case, there is a missing link for in vitro life cycle of adult worms and it is fertili-zation. Regarding the researchers’ observations, self-fertilization can be done in worms living in dog intestine, but despite all sorts of experimental techniques, this phenomenon has never been observed in reared worms in culture media. Further-more cross fertilization has not been observed in vitro and even in parasites with dog intestinal origin; although it theoretically is possible. During a follow-up of cultivated adult worms, evidences of behaviors similar to self-mating (Type 2) and cross-mating were observed in our lab which will be presented here.
Methods: Protoscoleces were aseptically removed from sheep hydatid cysts, washed twice with PBS and then cultivated in S.10E.H culture medium. The stages of parasite growth were observed using an inverted microscope for two months and all stages and behaviors were microscopically photographed. Different movies have also been made from these behavioral features.
Results: After around 55 days post cultivation, some evidences of behaviors simi-lar to self-mating (Type 2) and cross-mating were observed in some of the mature adult worms. However, fertile eggs in these parasites have never been observed.
Conclusion: Regarding the above observations, these parasites show tendency to unsuccessful self-mating/fertilization (type 2) which failure could be due to ana-tomical position and physiological maturation. Also lack of suitable conditions for self-fertilization causes the worms try to do unsuccessful cross- mating/fertilization in culture media.
Budke CM, Deplazes P, Torgerson PR. Global socioeconomic impact of cystic echinococcosis. Emerg Infect Dis. 2006; 12(2): 296-303.
Sadjjadi SM. Present situation of echinococcosis in the Middle East and Arabic North Africa. Parasitol Int. 2006; 55 Suppl: S197-202
Rokni MB. Echinococcosis / hyda-tidosis in Iran. Iran J Parasitol 2009; 4(2): 1-16.
Fasihi Harandi M, Budke CM, Ros-tami S. The monetary burden of cystic echinococcosis in Iran. PLoS Negl Trop Dis. 2012; 6(11): e1915.
Smyth JD. Studies on tapeworm physiology. XI. In vitro cultivation of Echinococcus granulosus from the protoscolex to the strobilate stage. Parasitology. 1967; 57(1): 111-133.
Smyth JD, Davies Z. In vitro culture of the strobilar stage of Echinococcus granulosus (sheep strain): A review of basic problems and results. Int J Par-asitol. 1974; 4: 631-644.
Smyth JD. Echinococcus granulosus and E. multilocularis: in vitro culture of the strobilar stages from protoscoleces. Angew Parasitol. 1979; 20(3): 137-147.
Smyth JD, Smyth MM. Self-insemination in E. granulosus in vivo. J Helminthol. 1969 ;43:383-388.
Kumaratilake LM, Thompson RC, Eckert J, D'Alessandro A. Sperm transfer in Echinococcus (Cestoda: Taeniidae). Z Parasitenkd. 1986; 72(2): 265-269.
Lymbery AJ, Thompson RC. Elec-trophoretic analysis of genetic varia-tion in Echinococcus granulosus from domestic hosts in Australia. Int J Parasitol. 1988; 18(6):803-811.
Haag KL, Araújo AM, Gottstein B, Siles-Lucas M, Thompson RC, Zaha A. Breeding systems in Echinococcus granulosus (Cestoda; Taeniidae): self-ing or outcrossing? Parasitology. 1999; 118 ( Pt 1): 63-71.
Mohammadzadeh T, Sadjjadi SM, Rahimi HR, Shams S. Establishment of a modified in vitro cultivation of protoscoleces to adult Echinococcus granulosus; an important way for new investigations on hydatidosis. Iran J Parasitol. 2012; 7(1): 55-66.
Smyth JD. In vitro cultivation of parasitic helminths. CRC press.1990; Pp: 123-137.
Dévé F. Evolution vesicular du sco-lex echinococcique obtenue in vitro. La culture artificielle de kyste hyda-tique. Comptes Rendus de la Société Biologique. 1926; 94: 440-450.
Dévé F. Scoliciculture hydatique en sac de collodion et in vitro. Comptes Rendus de la Société Biologique. 1928; 98: 1176-1177.
Smyth JD. Studies on tapeworm physiology, X. Axenic cultivation of the hydatid organism. Echinococcus granulosus; establishment of a basic technique. Parasitology .1962; 52(3-4): 441-457.
Smyth JD, Howkins AB. An in vitro technique for the production of eggs of Echinococcus granulosus by matura-tion of partly developed strobila. Parasitology. 1966; 56(4)763-766.
Smyth JD, Barrett NJ. Procedures for testing the viability of human hy-datid cysts following surgical remov-al, especially after chemotherapy. Trans R Soc Trop Med Hyg.1980; 74(5):649-652.
Macpherson CN, Smyth JD. In vitro culture of the strobilar stage of Echi-nococcus granulosus from protoscoleces of human, camel, cattle, sheep and goat origin from Kenya and buffalo origin from India. In J Parasitol. 1985; 15(2): 137-140.
Hijjawi NS, Abdel-Hafez SK, Kam-hawi SA. Echinococcus granulosus: pos-sible formation of a shelled egg in vitro. 1992; 22(1): 117-118.
Bölükbaş Cs, Doğanay A. Echinococ-cus granulosus’un in vitro Kϋltϋrϋ. Tü-rkiye Parazitoloji Dergisi. 2008; 32(4): 360-365.
Thompson RCA. Biology and sys-tematics of Echinococcus in: Thomp-son RCA, Lymbery AJ. Echinococcus and hydatid disease. Wallingford. CAB International. 1995. pp: 1-37.
Maillard S, Benchikh-Elfegoun MC , Kohil K, Gottstein B, Piarroux R. Failure to observe cross-fertilization between the Echinococcus granulosus G1 and G6 strains after an experi-mental mixed infection of the defini-tive host. Vet Parasitol. 2011; 175(1-2): 80–83.
Lymbery AJ, Hobbs RP, Thompson RC. The dispersion of Echinococcus granulosus in the intestine of dogs. J Parasitol. 1989; 75(4): 562-570.
Howell MJ, Smyth JD. Maintenance and cultivation of Echinicoccus species in vivo and in vitro in: Thompson RCA, Lymbery AJ. Echinococcus and hydatid disease Wallingford. CAB International. 1995.pp: 201-232.
Files | ||
Issue | Vol 9 No 1 (2014) | |
Section | Articles | |
Keywords | ||
Cross-mating/fertilization Culture media Echinococcus granulosus Self-mating/fertilization |
Rights and permissions | |
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |