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.
Identifer | oai:union.ndltd.org:NSYSU/oai:NSYSU:etd-0804106-212624 |
Date | 04 August 2006 |
Creators | Yang, Chu-Ya |
Contributors | Jung-Pin Hsu, Cheng-Fang Chang, Li-Lian Liu, Shih-Chu Chen |
Publisher | NSYSU |
Source Sets | NSYSU Electronic Thesis and Dissertation Archive |
Language | Cholon |
Detected Language | English |
Type | text |
Format | application/pdf |
Source | http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0804106-212624 |
Rights | withheld, Copyright information available at source archive |
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