Decreasing egg quality following oocyte ageing is a major restricting factor for the breeding programs. The mechanisms behind this process has not yet been clarified. To examine the possible involvement of oxidative stress in the oocyte ageing process, the relative mRNA abundance of specific transcripts were determined in oocytes collected from 6 females and incubated in vitro for 18 hours post stripping at 20 °C in goldfish Carassius auratus. During the 18 hour-post-stripping ageing of the oocytes, relative mRNA levels of candidate transcripts involved in oxidative injury, mitochondrial function and stress response, cell cycles, apoptosis, reproduction and germ line speciation and developmental competence were measured by real-time PCR. None of the relative mRNA abundance of the examined genes were significantly altered through oocyte ageing. In addition, the amount of thiobarbituric acid reactive substances (TBARS), an indicator of lipid peroxidation, did not change over time following stripping. The activity of the antioxidant enzymes also remained constant during oocyte ageing. The results of the current study indicated that oxidative stress unlikely plays a role as an initiator or promotor in the progress of oocyte ageing in goldfish.

• MeSH
• embryo nesavčí metabolismus   patologie   MeSH
• karas zlatý MeSH
• messenger RNA genetika   metabolismus   MeSH
• oocyty metabolismus   patologie   MeSH
• oxidace-redukce MeSH
• oxidační stres * MeSH
• peroxidace lipidů * MeSH
• rybí proteiny genetika   metabolismus   MeSH
• stanovení celkové genové exprese MeSH
• stárnutí metabolismus   patologie   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The spawning behavior of different fish species is as diverse as their habitats. A lot of factors influence the (phospho)lipid composition of fish sperm, including the water temperature at which spawning takes place. Therefore, this study aimed on the elucidation of the phospholipid composition of sperm from three fish species from different orders (common carp - Cyprinus carpio, northern pike - Esox lucius and burbot - Lota lota) with different spawning temperatures by nuclear magnetic resonance spectroscopy (NMR), matrix-assisted laser desorption/ionization mass spectrometry and thin-layer chromatography (TLC) coupled to electrospray ionization mass spectrometry as well as gas chromatography. Next to the lipid composition that was different for carp, northern pike and burbot, regarding the moieties of the different (phospho)lipid classes (particularly sphingomyelin and acidic phospholipids) and the saturation degree of the fatty acyl residues, there were differences observed depending on the analytical method that was used. The results from TLC and NMR investigations differed regarding the amounts of the different phospholipids. Reasons for these discrepancies are discussed in detail.

• MeSH
• druhová specificita MeSH
• lipidy chemie   MeSH
• magnetická rezonanční spektroskopie MeSH
• plavání * MeSH
• ryby fyziologie   MeSH
• sladká voda chemie   MeSH
• spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
• spermie chemie   fyziologie   MeSH
• teplota * MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

All extant groups of Elasmobranches have internal fertilization and the structure of the male reproductive organs is very specific: sperm passes from the internal organs via the cloaca, but the male copulating organ (clasper) is distant from the cloaca. This suggests that sperm can contact the surrounding medium before fertilization. Because of this involvement with the environment, external signaling in sperm motility activation could occur in these species even though their fertilization mode is internal. In this case, spermatozoa of Elasmobranches should hypothetically possess a specific structure and membrane lipid composition which supports physiological functions of the sperm associated with environmental tonicity changes occurring at fertilization. Additionally, sperm motility properties in these taxa are poorly understood. The current study examined sperm lipid composition and motility under different environmental conditions for the ocellate river stingray, Potamotrygon motoro, an endemic South America freshwater species. Sperm samples were collected from six mature males during the natural spawning period. Sperm motility was examined in seminal fluid and fresh water by light video microscopy. Helical flagellar motion was observed in seminal fluid and resulted in spermatozoon progression; however, when diluted in fresh water, spermatozoa were immotile and had compromised structure. Lipid class and fatty acid (FA) composition of spermatozoa was analyzed by thin layer and gas chromatography. Spermatozoa FAs consisted of 33 ± 1% saturated FAs, 28 ± 1% monounsaturated FAs (MUFAs), and 41 ± 1% polyunsaturated FAs (PUFAs), and a high content of n-6 FAs (32 ± 2%) was measured. These results allowed us to conclude that sperm transfer from P. motoro male into female should occur without coming into contact with the hypotonic environment so as to preserve potent motility. In addition, this unusual reproductive strategy is associated with specific spermatozoa structure and lipid composition. Low level of docosahexaenoic acid and relatively low PUFA/MUFA ratio probably account for the relatively low fluidity of freshwater stingray membrane and can be the main reason for its low tolerance to hypotonicity.

• MeSH
• analýza spermatu veterinární   MeSH
• lipidy chemie   MeSH
• motilita spermií fyziologie   MeSH
• rejnokovití fyziologie   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Oocyte ageing is the most important factor affecting egg quality of several fish species after ovulation. Oxidative stress has been proposed as the initiator of the oocyte ageing process in other vertebrates. To identify the role of oxidative stress and apoptosis on the progress of oocyte ageing in the common carp Cyprinus carpio, changes in the relative mRNA abundance of selected transcripts were examined. The possible alteration in the oxidation status of the oocytes during ageing was also studied. In addition, the activity of antioxidant enzymes during oocyte ageing was evaluated. Oocytes from 6 females were incubated in vivo for 14 hours post-ovulation (HPO) and in vitro for 10 hours post-stripping (HPS) at 20°C before fertilization. Hatching rates were over 65% up to 4-6 HPO, finally dropping to 1.3% at 12-14 HPO.Hatching rates were over 65% up to 4-6 HPO, finally dropping to 1.3% at 12-14 HPO. Hatching rates were more than 70% for the eggs stored in vitro up to 6 HPS and then decreased to 21.3% at 10 HPS. The results demonstrated no significant changes in the relative mRNA levels of oxidative stress-related genes or genes involved in the cell cycle during the progress of oocyte ageing in common carp. Additionally, the amount of TBARS and carbonyls did not change as time elapsed following ovulation. The apoptosis-related genes however, were significantly altered following the prolonged time interval between ovulation and fertilization. The lack of response of both activities of antioxidant enzymes and oxidation products during oocyte ageing strengthens the conclusion that oxidative stress is unlikely to be a main factor determining the progress of oocyte ageing in common carp. However, an increase in the mRNA abundance of apoptosis-related genes demonstrates that apoptotic pathway might be involved in the progress of oocyte ageing.

• MeSH
• antioxidancia metabolismus   MeSH
• kapři metabolismus   MeSH
• messenger RNA metabolismus   MeSH
• oocyty cytologie   metabolismus   MeSH
• oxidace-redukce MeSH
• oxidoreduktasy metabolismus   MeSH
• rybí proteiny metabolismus   MeSH
• stárnutí buněk * MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

This study investigates the replacement of vegetable oil (VO) in aquaculture feed for Arctic char (Salvelinus alpinus) with oil produced by the oleaginous yeast Lipomyces starkeyi grown in lignocellulose (wheat straw) hydrolysate. VO is extensively used to partially replace fish oil in aquaculture feed, which can be seen as non-sustainable. VO itself is becoming a limited resource. Plant oils are used in many different applications, including food, feed and biodiesel. Its replacement in non-food applications is desirable. For this purpose, yeast cells containing 43% lipids per g dry weight were mechanically disrupted and incorporated into the fish feed. There were no significant differences in this pilot study, regarding weight and length gain, feed conversion ratio, specific growth rate, condition factor and hepatosomatic index between the control and the yeast oil fed group. Fatty and amino acid composition of diet from both groups was comparable. Our results in fish demonstrate that it is possible to replace VO by yeast oil produced from lignocellulose, which may broaden the range of raw materials for food production and add value to residual products of agriculture and forestry.

• MeSH
• aminokyseliny analýza   MeSH
• krmivo pro zvířata analýza   MeSH
• lignin metabolismus   MeSH
• Lipomyces růst a vývoj   metabolismus   MeSH
• mastné kyseliny analýza   chemie   MeSH
• pilotní projekty MeSH
• pšenice metabolismus   MeSH
• pstruh růst a vývoj   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The aim of this study was to compare the spermatozoa quality parameters in spermatozoa of RAS (Recirculating Aquaculture System; RAS group) cultured (commercial pellets) and natural condition cultured (WILD group) burbot Lota lota (live prey, Pseudorasbora parva). Seven of nine fish of the RAS group produced sperm, with sperm from only four of the fish having a motility of >5%. Sperm were collected from all nine fish of the WILD group, and sperm of six of the fish from the WILD group had motility of about 100% and three had sperm with 50% to 60% motility. Spermatozoa from the RAS group had a delay in activation compared to the WILD group. Fish from the RAS group also had a lesser volume of sperm (1.8 ± 1.2 mL) collected compared to the WILD group (3.6 ± 1.2 mL). Compared to the RAS group, sperm of the WILD group had a greater proportion of saturated fatty acids (SFA), as well as the phospholipid, phosphatidylethanolamine. The findings indicate that fish grown in natural conditions may be more suitable as broodstock. Ongoing research to develop methods of enhancing reproductive performance of burbot broodstock cultured in RAS is needed to investigate whether the quality of sperm can be improved by adjusting environmental conditions, diet, or combination of these factors.

• MeSH
• analýza spermatu * MeSH
• divoká zvířata MeSH
• Gadiformes metabolismus   fyziologie   MeSH
• lipidy analýza   MeSH
• mastné kyseliny analýza   metabolismus   MeSH
• metabolismus lipidů * MeSH
• spermie chemie   metabolismus   MeSH
• vodní hospodářství metody   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

The intake of cadmium contaminated fish was mimicked by incubating human hepatoblastoma cells (Cell line HepG2) with a combination of different levels of cadmium (0-5μM) plus the n-3 fatty acids docosahexaenoic acid and eicosapentaenoic acid, which are typical for fish. Uptake of cadmium, iron, copper and zinc was measured by ICP-MS. In addition mRNA expression of two metallothioneins (mt1 g and mt1 m) was evaluated by real-time PCR. The obtained data shows that the presence of cadmium increases the uptake of iron and zinc into the HepG2 cells while the uptake of copper remains unaffected. The presence of the chosen fatty acids did not affect the uptake of either cadmium or iron, zinc and copper. The presence of already 1μM cadmium increased the mRNA expression of mt1 g and mt1 m significantly, while the fatty acids did not interfere with the effect of cadmium.

• MeSH
• biologický transport účinky léků   MeSH
• buňky Hep G2 MeSH
• exprese genu účinky léků   MeSH
• kyselina eikosapentaenová farmakologie   MeSH
• kyseliny dokosahexaenové farmakologie   MeSH
• lidé MeSH
• messenger RNA metabolismus   MeSH
• metalothionein genetika   MeSH
• těžké kovy farmakologie   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

In fish, sperm quality is frequently associated with sperm motility variables. The response of sperm motility to different temperatures varies among species and plasma membrane lipid composition may contribute to variations in findings in previous research. In the present study, sperm motility and lipid composition were analysed between motile or immotile carp Cyprinus carpio sperm at different in vitro temperatures (4, 14 and 24°C). The duration of the period over which sperm motility is sustained was longer at 4°C compared with 14 and 24°C; while sperm velocity was greatest at 24°C. Motile sperm had lesser proportions of 18:3 (n-3) and 22:6 (n-3) fatty acids at 24°C relative to immotile sperm. There was no difference in fatty acid composition of motile and immotile sperm at 4 and 14°C. The total phospholipid content was less in motile than in immotile sperm at 24°C. At 24°C, phosphatidylcholine and phosphatidylserine proportions were less in motile than immotile sperm. It is concluded that lipid composition of motile carp sperm is affected by temperature, with greater temperatures associated with reduced lipid content, elevation of sperm curvilinear velocity and a decreased duration of the period over which motility is sustained.

• MeSH
• kapři fyziologie   MeSH
• lipidy chemie   MeSH
• motilita spermií fyziologie   MeSH
• spermie chemie   fyziologie   MeSH
• teplota * MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: Human hepatocellular cells Hep G2 were used to mimic and investigate the effect of the intake of cadmium (Cd(2+)) contaminated fish on cytotoxicity, fatty acid (FA) and phospholipid class composition. METHODS: Cells were incubated with a combination of Cd(2+) and eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) fish specific FA. RESULTS: We measured a significant increased proportion of EPA and DHA in the treated cells compared to the control line confirming the uptake. While doses of 25 μM DHA showed to be toxic to the cells, repeated short term incubations (2 h) at lower doses resulted in an increased uptake of DHA. The resarzurin assay, evaluating cell viability, showed a significant decrease in cell viability between Cd(2+) incubation time and, further, the pre-incubation with BSA-FA complex resulted in significantly increased cell viability. On the metabolic level, increased concentrations of EPA and DHA resulted in an increased proportion of docosapentaenoic acid (DPA) which indicated an increased metabolism. Also 24-h combined incubations of 5 μM Cd(2+) and EPA and DHA showed a significant increase DPA in the total lipid fraction of the cells. In addition, incubation with 5 μM Cd(2+) for 24 h also decreased the total cardiolipin (CL) fraction from the identified phospholipids. CONCLUSIONS: We confirmed that the applied FA were taken up by the cells. A combination of EPA, DHA and Cd(2+) influenced lysosomal integrity, cell viability and lipid metabolism in the cells. The most important finding is that DHA and EPA reduced the detrimental effect of Cd(2+) on cell viability. The exact effects and kinetics behind our observations still need further evaluation.

• MeSH
• bezpečnost potravin MeSH
• buňky Hep G2 MeSH
• chemické látky znečišťující vodu metabolismus   toxicita   MeSH
• fyziologická absorpce MeSH
• hepatocyty účinky léků   metabolismus   MeSH
• kadmium metabolismus   toxicita   MeSH
• kyselina eikosapentaenová metabolismus   MeSH
• kyseliny dokosahexaenové metabolismus   MeSH
• lidé MeSH
• metabolismus lipidů MeSH
• nenasycené mastné kyseliny metabolismus   MeSH
• potrava z moře (živočišná) MeSH
• ryby metabolismus   MeSH
• viabilita buněk MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH