In this project, the eggs of tench (Tinca tinca) were used for the evaluation of changes in fatty acid (FA) composition and lipid classes during in vitro ova ageing until the occurrence of over-ripening. Stripped ova of 6 females were collected separately and stored in sterile cell culture plates in an incubator at 20 °C for up to 24 hours post stripping (HPS). Stored ova were fertilized at 0, 2, 4, 6, 8, 10 and 24 HPS. Egg eyeing and hatching rates were assessed as indices of egg quality. Upon fertilization at each HPS, samples were taken separately frozen in liquid nitrogen and stored at -80°C till further analyses. Differences in the FA and lipid composition and embryo survival rates at the different fertilization times post-stripping were evaluated. The lipid fraction was extracted from the oocytes according to Hara and Radin (1978). The lipid content of the samples was determined gravimetrically from total extracted lipid. The FA were then analysed with a gas chromatograph equipped with a flame ionisation detector and PVT injector. The peaks were identified by comparing their retention times with those of authentic standards. Major lipid classes and the phospholipid classes were separated according to (Olsen and Henderson, 1989). Quantitative analysis of the separated lipid- and phospholipid classes was done by scanning the plates after derivatisation with a CAMAG TLC Scanner 3 (Camag, Switzerland). Identification of the lipid classes was performed by comparison with authentic standards applied on the same plate. In present study no significant changes were observed in lipid content and composition during storage of eggs until over-ripening. As well FA composition did not change over time indicating another reason of deterioration of oocyte quality than the proposed lipid oxidation. However individual fishes in this study showed much different fertilization rates that corresponded with their levels of cholesterol and triacylglycerols in minor degree.
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:320763 |
| Date | January 2017 |
| Creators | KRZYŚKÓW, Anna |
| Source Sets | Czech ETDs |
| Language | English |
| Detected Language | English |
| Type | info:eu-repo/semantics/masterThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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