첨가제의 종류와 동해방지제의 농도에 따른 피조개, Scapharca broughtonii D형 유생의 냉동보존 효과 비교

  • 조필규 (부경대학교 수산과학대학 양식학과) ;
  • 최윤희 (부경대학교 수산과학대학 양식학과) ;
  • 강경호 (여수대학교 수산생명공학부) ;
  • 고강희 (여수대학교 수산생명공학부) ;
  • 고창순 (국립수산과학원 남해수산종묘시험장) ;
  • 김병학 (국립수산과학원 남해수산종묘시험장) ;
  • 임한규 (국립수산과학원 울진수산종묘시험장) ;
  • 최철영 (미국 국립보건원) ;
  • 장영진 (부경대학교 수산과학대학 양식학과)
  • Jo, Pil-Gue (Department of Aquaculture, Pukyong National University) ;
  • Choi, Youn-Hee (Department of Aquaculture, Pukyong National University) ;
  • Kang, Kyoung-Ho (Department of Aquaculture, Yosu National University) ;
  • Kho, Kang-Hee (Department of Aquaculture, Yosu National University) ;
  • Go, Chang-Soon (Namhae Marine Hatchery, National Fisheries Research and Development Institute) ;
  • Kim, Byong-Hak (Namhae Marine Hatchery, National Fisheries Research and Development Institute) ;
  • Lim, Han-Kyu (UlJin Marine Hatchery, National Fisheries Research and Development Institute) ;
  • Choi, Cheol-Young (Laboratory of Gene Regulation and Development, National Institute of Child Health and Human Development, National Institutes of Health(NIH)) ;
  • Chang, Young-Jin (Department of Aquaculture, Pukyong National University)
  • 발행 : 2002.12.01

초록

피조개 D형 유생을 냉동보존하기 위한 적절한 첨가제 종류와 농도 및 동해방지제로 적용한 ethylene glycol 농도에 대한 냉동보존 효과실험을 실시하였다. 유생을 0$^{\circ}C$로 설정된 프로그램 냉동기로 옮겨 -1$^{\circ}C$/분의 냉동율로 -12$^{\circ}C$까지 냉동한 후 10분 동안 유지한 상태에서 식빙하고 -35$^{\circ}C$까지 재냉동한 다음, 즉시 액체질소통에 저장하였다. 1.0 M과 2.0 M ethylene glycol을 동해방지제로 사용하여 첨가제별로 D형 유생을 냉동한 결과, 0.5 M sucrose에서 생존율이 각각 50.5 ${\pm}$ 1.3%, 51.9 ${\pm}$ 1.7%로 가장 좋았다. 0.5 M sucrose를 첨가한 ethylene glycol의 농도별 냉동에서는 2.0 M이 생존율 51.9 ${\pm}$ 1.7%로 가장 높았다. 주사 및 투과전자현미경을 이용하여 유생을 관찰한 결과, 냉동 전, 후 유생의 외부형태 및 세포내 구조의 차이가 인정되지 않았다.

This study was performed to find out the optimal kind of additive and concentration of ethylene glycol (EG) as the cryoprotectant on the D-shaped larvae of arkshell, Scapharca broughtonii. The larvae in straws was carried in the programed freezer set uo at 0$^{\circ}C$, frozen to -12$^{\circ}C$ by the freezing rate of -1$^{\circ}C$/min, held for 10 minutes, seeded at -12$^{\circ}C$, finally refrozen to -35$^{\circ}C$ by the freezing rate of -1$^{\circ}C$/min and then directly plunged into liquid nitrogen. The survival of larvae frozen in 1.0 M and 2.0 M ethylene glycol added with 0.5 M sucrose were high (50.5 ${\pm}$ 1.3% and 51.9 ${\pm}$ 1.7%, respectively) in the early D-shaped larvae cryopreserved in 1 M and 2 M ethylene glycol diluted with 0.2 M and 0.5 M fructose, glucose and sucrose. In the late D-shaped larvae were cryopreserved according to five concentrations of ethylene glycol added with 0.5 M sucrose, the survival of larvae frozen in 2.0 M ethylene glycol was the highest as 51.9 ${\pm}$ 1.7%. The morphological differences in cells between unfrozen and frozen-thawed D-shaped larvae were not found by the scanning and transmission electron microscopy.

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