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Studies on the cryopreservation and in vitro culture of Amyloodinium ocellatum

The Amyloodinium ocellatum was collected from cobia ( Rachycentron canadum ) gill and four tests including 4 ¢J storage, toxicity of cryoprotectant, cryopreservation and in vitro cultivation on fish cell line were conducted to establish the methods of preservation of Amyloodinium ocellatum. Survival of trophont, morphology and division of tomont and number of dinospore released were evaluated the effects of this study. The results showed that division irregulated, delayed and stopped of the tomont were found after stored at 4 ¢J over 48 hours. It was produced 1.08 x 10 4 cell/ml dinospores from 1 x 10 3 trophont at 4 ¢J, 24 hours storage group and significant higher ( p¡Õ0.0001 ) than other storage groups. For the toxicity of cryoprotectant, the concentration of DMSO 3~10¢M, Glycerol 3~10¢M, Methanol 3~10¢M, Ethanol 3~5¢M, PrOH 3~5¢M, DMAc 3~5¢M, Sucrose 3~15¢M, Trehalose 3~15¢M, Dextran 3~5¢Mand Ficoll 3~10¢Mwere safety to use on A. ocellatum trophont preservation. It was unsuccessful to cryopreserve the trophont of A. ocellatum when stored at direct liquid N2 freezing, different -20 ¢J freezing time, -1 ¢J min-1 freezing container and different cryoprotectant equilibration time contain 10¢MGlycerol and DMSO, respectively. Using the U-shaped tube of sigle and double loop could gain pure and bacteria-free dinospores. The results of in vitro cultivation of A. ocellatum showed that eel epidermis and cobia fin cell line with different culture mediums were unable to grow the trophont and tomont of A. ocellatum.

Identiferoai:union.ndltd.org:NSYSU/oai:NSYSU:etd-0804106-212624
Date04 August 2006
CreatorsYang, Chu-Ya
ContributorsJung-Pin Hsu, Cheng-Fang Chang, Li-Lian Liu, Shih-Chu Chen
PublisherNSYSU
Source SetsNSYSU Electronic Thesis and Dissertation Archive
LanguageCholon
Detected LanguageEnglish
Typetext
Formatapplication/pdf
Sourcehttp://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0804106-212624
Rightswithheld, Copyright information available at source archive

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