Stereological comparison of oocyte recruitment and batch fecundity estimates from paraffin and resin sections using spawning albacore (Thunnus alalunga) ovaries as a case study.

Traditional histological protocols in marine fi sh reproductive laboratories using paraf fi n as the embedding medium are now increasingly being replaced with protocols using resin instead. These procedures entail differ- ent degrees of tissue shrinkage complicating direct comparisons of measurement results across laboratories or articles. In this work we selected ovaries of spawning Mediterranean albacore ( Thunnus alalunga ) as the subject of our study to address the issue of structural changes, by contrasting values on oocyte recruitment and fi nal batch fecundity given from the same tissue samples in both paraf fi nandresin.Amodernstereologicalmethod, the oocyte packing density (OPD) theory, was used supported by initial studies on ovarian tissue sampling and measurement design. Examples of differences in the volume fraction of oocyte stages, free space and connective tissue were found between the embedding media. Mean oocyte diameters were smaller in paraf fi nthaninresin with differences ranging between 0.5% in primary growth and 24.3% in hydration (HYD) stage oocytes. Fresh oocyte measurements showed that oocytes shrank as a consequence of the embedding process, reaching the maximaldegreeofshrinkageforoocytesintheHYDstage(45.8%inparaf fi nand26.5%inresin).Inordertoassess the effect of oocyte shrinkage on the OPD result, and thereby on relative batch fecundity (F r ), oocyte diameters corrected and uncorrected for shrinkage, were used for estimations. Statistical signi fi cant differences were found ( P b 0.05) between these two approaches in both embedding media. The average F r was numerically smaller in paraf fi n compared to resin (86 ± 61 vs. 106 ± 54 oocytes per gram of body mass (mean ± SD)). For both embedding media statistical signi fi cant differences ( P b 0.05) were seen between F r results based on either oocytes in the germinal vesicle migration stage or HYD stage. As a valuable adjunct, the present use of the OPD theory made it possible to document that the oocyte recruitment of spawning ovaries of Mediterranean albacore followed the typical pattern of an asynchronous oocyte development and indeterminate fecundityPostprint2,444

Demonstrations of maternal effects of Atlantic cod: combining the use of unique mesocosm and novel molecular techniques - a new EU-project

One of the effects of fishing is a reduction in average age and size in exploited fish stocks, leading to an increasing proportion of recruit spawners in the stock. Current management practice assumes equal viability of offspring from first time spawners and from repeat spawners, despite the fact that that first time spawners often produce smaller eggs than older spawners. The aim of this EU-project is to follow offspring from families of first time spawning and older cod, reared under identical and semi-natural conditions in marine enclosures (mesocosms). The parental origin of the larvae is identified using microsatellite DNA methodology. The advantage of this approach, compared to traditional laboratory rearing, is that rearing conditions are close to natural conditions, and all larvae are reared in the same environment. This eliminates the tank-to-tank variability often observed in traditional rearing experiments. The fish are reared from hatching, through the larval and juvenile stages, until sexual maturity. Growth rates, survival and nutritional condition will be measured using methods such as RNA/DNA ratio and otolith micro increment analysis. The results will be related to parental origin and quality measures of the eggs. It is intended to incorporate the results into management models for improvement of fishery management strategies. In this paper we will focus on a description of the projec

Egg production of Baltic cod (Gadus morhua) in relation to variable sex ratio, maturity, and fecundity

Observed fluctuations in relative fecundity of Eastern Baltic cod (Gadus morhua L.) were related to food availability during the main feeding period and were used to develop a predictive model that explained 72% of the interannual variations in fecundity. Time series of sex ratios, maturity ogives, and relative fecundity were combined with mean weights-at-age and stock sizes from an analytical multispecies model to estimate the potential egg production (PEP). Relationships between PEP and independent estimates of realized daily and seasonal egg production from egg surveys were highly significant. The difference between estimates of potential and realized seasonal egg production was of a magnitude corresponding to the expected loss of eggs as a result of atresia, fertilization failure, and early egg mortality. The removal of interannual variability in sex ratio, maturity, and fecundity on estimates of PEP deteriorated the relationships in all three cases. PEP proved to be superior to spawning stock biomass as measure of the reproductive potential in a stock-recruitment relationship of Eastern Baltic cod. PEP in combination with the reproductive volume explained 61% of the variation in year-class strength at age 2

Egg quality determinants in cod (Gadus morhua L.): egg performance and lipids in eggs from farmed and wild broodstock

Lipids and essential fatty acids, particularly the highly unsaturated fatty acids, 20:5n-3 (eicosapentaenoic acid; EPA), 22:6n-3 (docosahexaenoic acid; DHA) and 20:4n-6 (arachidonic acid, AA) have been shown to be crucial determinants of marine fish reproduction directly affecting fecundity, egg quality, hatching success, larval malformation and pigmentation. In Atlantic cod (Gadus morhua L.) culture, eggs from farmed broodstock can have much lower fertilisation and hatching rates than eggs from wild broodstock. The present study aimed to test the hypothesis that potential quality and performance differences between eggs from different cod broodstock would be reflected in differences in lipid and fatty acid composition. Thus eggs were obtained from three broodstock, farmed, wild/fed and wild/unfed, and lipid content, lipid class composition, fatty acid composition and pigment content were determined and related to performance parameters including fertilisation rate, symmetry of cell division and survival to hatching. Eggs from farmed broodstock showed significantly lower fertilisation rates, cell symmetry and survival to hatching rates than eggs from wild broodstock. There were no differences in total lipid content or the proportions of the major lipid classes between eggs from the different broodstock. However, eggs from farmed broodstock were characterised by having significantly lower levels of some quantitatively minor phospholipid classes, particularly phosphatidylinositol. There were no differences between eggs from farmed and wild broodstock in the proportions of saturated, monounsaturated and total polyunsaturated fatty acids. The DHA content was also similar. However, eggs from farmed broodstock had significantly lower levels of AA, and consequently significantly higher EPA/AA ratios than eggs from wild broodstock. Total pigment and astaxanthin levels were significantly higher in eggs from wild broodstock. Therefore, the levels of AA and phosphatidylinositol, the predominant AA-containing lipid class, and egg pigment content were positively related to egg quality or performance parameters such as fertilisation and hatching success rates, and cell symmetry

Thermal dynamics of ovarian maturation in Atlantic cod (Gadus Morhua)

The timing and success of spawning in marine fish are of fundamental importance to population persistence and distribution and, for commercial species, sustainability. Their physiological processes of reproduction are regulated, in part, by water temperature, and therefore changes in marine climate may have dramatic effects on spawning performance. Using adult Atlantic cod (Gadus morhua) as a case study, we examined the links between water temperature, body size, vitellogenesis, and spawning time by conducting extensive laboratory and field studies. Our experiments documented that vitellogenesis generally starts at autumnal equinox and that oocyte growth and investment are greater in cod held at warmer temperatures. Furthermore, spawning occurred earlier when oocyte growth was more rapid. Large females spawned earlier than smaller females at warmer temperatures, but this effect vanished at colder temperatures. The experimental results were confirmed by measurements of oocyte growth collected from wild-caught cod in northern (Barents Sea) and southern (Irish Sea and North Sea) populations. The established, general model of oocyte maturation was consistent with published egg production curves of cod from these waters, considering relevant in situ temperatures recorded by individual data-storage tags on cod. These findings have considerable relevance for future studies of fish recruitment in relation to climate change