| In vitro cultivation for development of Ascaris suum from the decoated and embryonated eggs(second-stage larva) |
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Cha-ho Jee, Seung-jun Park |
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College of Veterinary Medicine, Chungbuk National University |
| 시험관내에서 돼지회충(Ascaris suum) 함자충란(L2)의 인공배양 |
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지차호, 박승준 |
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충북대학교 수의과대학 |
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| Abstract |
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The cultivation for development of Ascaris suum from the second-stage larvae($L_2$) embryonated egg and the third-stage of rat-derived larvae($L_3$) recovered from lung of rats were performed to use the screening test of anthelmintics in vitro. The preparations of larvae for cultivation were that the artificially-hatched $L_2$ incubated the embryonated eggs of Ascaris suum in 0.1% formalin solution at $25^{circ}C$ for 28 days and the rat-derived larvae($L_3$) recovered from the lung of rat infected with the embryonated eggs of Ascaris suum on 7 days after infection(DAI). The cultivation for development of Ascaris suum from the embryonated eggs($L_2$) and the rat-derived larvae($L_3$) for 14 days in RPMI medium 1640(with 5% bovine calf serum) were as follows : 1. The sizes of the liberated larvae($L_2$) which were artificially hatched from embryonated eggs with glass beads(diameter 5mm) were $190{sim}250{mu}m$ on 1 days in culture(DIC). The second-stage larvae were molted into third-stage larvae(early $L_3$; $250{sim}300{mu}m$) and the features of these larvae were first observed such as cephalic cuticle, esophageal lumen and anus etc. on 5 DIC and the sizes of late third-stage larvae were $250{sim}450{mu}m$ on 10 DIC. The sizes of early fourth-stage larvae($L_4$) were $500{sim}700{mu}m$ and the features of these larvae were more pronounced in internal organs on 15 DIC. 2. The sizes of third-stage larvae($L_3$) recovered from the lung of rats were $1,340{sim}1,370{mu}m$ and the feartures of cephalic cuticle, esophageal lumen, intestine, rectum, anus were visualized by inverted microscope on 1 DIC. The fourth-stage larva($L_4$) completed by third ecdysis were recognizable and sizes of early fourth-stage larvae were developed as $1,400{sim}2,200{mu}m$ on 5 DIC. The sizes of middle fourth-stage of larva were $1,900{sim}2,300{mu}m$ and the thickened epithelial rectum was observed on 10 DIC. The rectum and anus of late fourth-stage larva($L_4$ $2,500{sim}3,200{mu}m$) had developed completely in RPMI medium 1640 on 15 DIC. |
| Key Words:
In vitro cultivation, embryonated egg of Ascaris suum, artificial hatching, RPMI medium 1640 |
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