Description: The stable production of high-quality fry in marine aquaculture is still hampered by unpredictable mortality caused by infectious diseases during larval rearing. Consequently, the development of new biocontrol agents is crucial for a viable aquaculture industry. The bacterial energy storage compound poly-β-hydroxybutyrate (PHB) has been shown to exhibit beneficial properties on aquatic organisms such as enhanced survival, growth, disease resistance and a controlling effect on the gastrointestinal microbiota. However, the effect of PHB on the developing immune system of fish larvae has so far not been investigated. In this study, the effect of feeding PHB-enriched Artemia nauplii on survival, growth and immune response in European sea bass (Dicentrarchus labrax) postlarvae was examined. Amorphous PHB was administered to 28-day-old sea bass postlarvae over a period of 10 days. The survival and growth performance were monitored, and the expression of 29 genes involved in immunity, growth, metabolism and stress-response was measured. While the expression of the insulin-like growth factor 1 (igf1), an indicator of relative growth, was upregulated in response to feeding PHB, the larval survival and growth performance remained unaffected. After 10 days of PHB treatment, the expression of the antimicrobial peptides dicentracin (dic) and hepcidin (hep) as well as mhc class IIa and mhc class IIb was elevated in the PHB fed postlarvae. This indicates that PHB is capable of stimulating the immune system of fish early life stages, which may be the cause of the increased resistance to diseases and robustness observed in previous studies.

Description: Productivity of marine fish stocks is known to be affected by environmental and ecological drivers, and global climate change is anticipated to alter recruitment success of many stocks. While the direct effects of environmental drivers on fish early life stage survival can be quantified experimentally, indirect effects in marine ecosystems and the role of adaptation are still highly uncertain. We developed an integrative model for the effects of ocean warming and acidification on the early life stages of Atlantic cod in the Barents Sea, termed SCREI (Simulator of Cod Recruitment under Environmental Influences). Experimental results on temperature and CO2 effects on egg fertilization, egg and larval survival and development times are incorporated. Calibration using empirical time series of egg production, temperature, food and predator abundance reproduces age-0 recruitment over three decades. We project trajectories of recruitment success under different scenarios and quantify confidence limits based on variation in experiments. A publicly accessible web version of the SCREI model can be run under www.oceanchange.uni-bremen.de/;SCREI. Severe reductions in average age-0 recruitment success of Barents Sea cod are projected under uncompensated warming and acidification toward the middle to end of this century. Although high population stochasticity was found, considerable rates of evolutionary adaptation to acidification and shifts in organismal thermal windows would be needed to buffer impacts on recruitment. While increases in food availability may mitigate short-term impacts, an increase in egg production achieved by stock management could provide more long-term safety for cod recruitment success. The SCREI model provides a novel integration of multiple driver effects in different life stages and enables an estimation of uncertainty associated with interindividual and ecological variation. The model thus helps to advance toward an improved empirical foundation for quantifying climate change impacts on marine fish recruitment, relevant for ecosystem-based assessments of marine systems under climate change.

Description: The growth and development of the aragonitic CaCO3 otoliths of teleost fish could be vulnerable to processes resulting from ocean acidification. The potential effects of an increase in atmospheric CO2 on the calcification of the otoliths were investigated by rearing Atlantic cod Gadus morhua L. larvae in 3 pCO2 concentrations—control (370 µatm), medium (1800 µatm) and high (4200 µatm)—from March to May 2010. Increased otolith growth was observed in 7 to 46 d post hatch (dph) cod larvae at elevated pCO2 concentrations. The sagittae and lapilli were usually largest in the high pCO2 treatment followed by the medium and control treatments. The greatest difference in mean otolith surface area (normalized to fish length) was for sagittae at 11 dph, with medium and high treatments being 46 and 43% larger than the control group, respectively. There was no significant pCO2 effect on the shape of the otoliths nor were there any trends in the fluctuating asymmetry, defined as the difference between the right and left sides, in relation to the increase in otolith growth from elevated pCO2.

Description: Plankton sampling was conducted in the Baltic to obtain sprat larvae. Their individual drift patterns were back-calculated using a hydrodynamic model. The modelled positions along the individual drift trajectories were subsequently used to provide insight into the environmental conditions experienced by the larvae. Autocorrelation analysis revealed that successive otolith increment widths of individual larvae were not independent. Otolith increment width was then modelled using two different generalized additive model (GAM) analyses (with and without autocorrelation), using environmental variables determined for each modelled individual larval position as explanatory variables. The results indicate that otolith growth was not only influenced by the density of potential prey but was controlled by a number of simultaneously acting environmental factors. The final model, not considering autocorrelation, explained more than 80% of the variance of otolith growth, with larval age as a factor variable showing the strongest significant impact on otolith growth. Otolith growth was further explained by statistically significant ambient environmental factors such as temperature, bottom depth, prey density and turbulence. The GAM analysis, taking autocorrelation into account, explained almost 98% of the variability, with the previous otolith increment showing the strongest significant effect. Larval age as well as ambient temperature and prey abundance also had a significant effect. An alternative approach applied individual-based model (IBM) simulations on larval drift, feeding, growth and survival starting as exogenously feeding larvae at the backcalculated positions. The IBM results revealed optimal growth conditions for more than 97% of the larvae, with a tendency for our IBM to slightly overestimate larval growth.

Description: We examined the RNA and DNA concentration of field-caught scallops Chlamys islandica, maintained in suspended cultures at 15 and 30 m depth, and scallops from a wild population at 50 to 60 m in Kobbefjord, southwest Greenland. General relations between RNA and DNA concentrations and individual shell height were established, and we found that the RNA:DNA ratio (RD) worked well as a standardisation of the RNA concentration independent of size and sex. During an experimental period of 14 mo, we observed a pronounced seasonal pattern in RD and mass growth, and differences between depths. Even though the period with high levels of RD reflected the growth season relatively well, RD was a poor predictor of individual mass growth rates of C. islandica. However, we found a non-linear response in RD to increased food concentrations resulting in RD being up- and down-regulated at the beginning and end of the productive summer season, respectively. These results indicate that short-term dynamics in the actual mass growth rate might be controlled through regulation of ribosome activity rather than ribosome number (RNA concentration). This adaption would allow scallops to up-regulate protein synthesis more rapidly, thereby ensuring efficient utilisation of the intense peaks in food availability in coastal areas in the Arctic. Therefore, we suggest that RD in C. islandica reflects the growth potential rather than the actual growth rate. Still, the amount of unexplained variance in RD is considerable and not independent over time, suggesting the existence of unresolved mechanisms or relationships.

Description: Recent intensification efforts of astacid culture considerably depend on the appropriate assessment of the animal's physiological condition both for research and application. We conducted a 4 weeks feeding experiment to assess temporal resolution and accuracy of different response parameters (RNA/DNA ratio, RNA per wet weight, carapace length, wet weight, specific growth rate). Juvenile noble crayfish were exposed to five feeding regimes that differed in feeding frequency and food availability. Continuous growth was detected in all feeding regimes with individual increase up to 90% (wet weight) and 17% (carapace length), respectively. Morphometric parameters allowed separation of three weight-groups or two length-groups. During the experimental period RNA/DNA ratios showed both decrease (−17%) and increase (+35%), with superior accuracy than morphometric parameters, separating four groups. Based on RNA/DNA ratios, different feeding regimes were detected earlier, with two groups separated already after 3 weeks. RNA/DNA ratio was clearly superior to RNA per unit wet weight, as the latter failed to detect any differences between groups. In conclusion, RNA/DNA ratio is a valuable tool in nutritional studies with freshwater crayfish if overall growth is the key variable of interest.

Description: The RNA/DNA ratio commonly used as proxy for the nutritional condition of fish larvae is affected by RNA degradation during analysis. For evaluation of two strategies to improve RNA integrity, a three-week feeding trial was carried out to assess the suitability of two nematode species (fam. Panagrolaimidae) as feed for newly hatched carp larvae (Cyprinus carpio) in comparison to Artemia nauplii (Artemia sp.) and a commercial dry feed. Aiming for an increased reproducibility of RNA/DNA determination, a high-salt inactivation (RNA later) as well as a targeted approach with a recombinant RNase inhibitor were compared to the classical protocol using lab chip technology. Improved RNA integrity was observed with high-salt inactivation when compared with a strategy applying a specific RNase inhibitor or the classic protocol. Carp larvae fed Artemia for 2 weeks and then dry feed for 1 week revealed the best overall growth performance as well as survival [83.0 ± 35.2 mg fresh weight (FW), 20.0 ± 2.4 mm total length (TL), 86.6 ± 11.7% survival]. Larvae fed the nematode species Panagrellus redivivus for 1 week and subsequently dry feed for 2 weeks (37.4 ± 29.1 mg FW, 14.7 ± 2.8 mm TL, 76.0 ± 6.0% survival) performed better than larvae fed with dry feed alone (28.2 ± 29.6 mg FW, 14.3 ± 2.9 mm TL, 54.3 ± 14.2% survival) or those receiving Panagrellus for 2 weeks. Between both nematode species, Panagrellus was a better feed with regard to growth performance and survival. RNA/DNA ratios ranged between 0.65 ± 0.27 (8 days post-hatch) and 1.96 ± 0.63 (22 days post-hatch) and were in the same treatment order as the other growth parameters. RNA/DNA ratios were significantly correlated with the growth rate, and decreasing RNA/DNA ratios in larger larvae may reflect decreasing growth rates with size rather than decreased nutritional status. Here, an improved RNA/DNA ratio protocol is presented in a feeding trial that reveals the suitability of nematodes as a first feed for common carp larvae.

Description: We investigated the nutritional condition of larval fishes caught in daily ichthyoplankton hauls in the ‘Helgoland Roads’ (54°11.18’N and 07°54.00’E) from February to June 2004. We concentrated on larvae of dab Limanda limanda and lesser sandeel Ammodytes marinus in order to compare early life stages of iteroparous and nearly semelparous fish. We analysed length, weight and RNA:DNA ratios as a proxy for condition of the larvae. The relationship between larval nutritional condition and larval size provided an indication of condition-selective mortality due to a loss of larvae in poor condition from the larger size-classes. For the sandeel, larvae in good condition were present in all size-classes, whereas for the dab maximum larval condition increased with increasing size. Variability in both standard length and condition was high in the 2 species during their planktonic stage. Both species showed good nutritional condition in the early to mid portion of the recruitment period and declines in condition in late April. This was more pronounced in larval dab, which showed a higher dependency on feeding conditions than larval sandeel. Together, these findings indicate a more conservative strategy of early life stages of the nearly semelparous sandeel compared with the iteroparous dab.

Description: The RNA/DNA ratio is a useful indicator of the nutritional condition of fish larvae. The presented analytical procedure is an improvement of Clemmesen's (Meeresforschung 32: 134-143, 1988) methodology which involves purification of fish larvae tissue homogenates and subsequent fluorescence-photometric measurements using specific nucleic acid dyes. The modifications concern the homogenization and nucleic acid extraction procedures. A 'shaking mill' was compared to a potter Elvehjem microhomogenizer and a reduction in the washing and purification steps was achieved. Treatment of samples with ribonuclease A and subsequent fluorescence measurement using ethidium bromide was given preference compared to the DNA-bisbenzimidazole determinations due to problems arising from high self-fluorescence of the samples and the influence of 'quenching' substances disturbing the DNA-bisbenzmidazole determinations. Different RNase concentrations and their influences on RNA and DNA were checked. Recovery rates of standard RNA and DNA 'spikes' were determined. Fish larvae samples were analysed with the previous and the improved modified procedure and a correction factor to compare results measured with the 2 procedures was calculated. With the presented method the physiological condition of individual larvae and the amount of variability can be determined.

Description: Somatic growth and nucleic acid content were studied in North Sea houting Coregonus oxyrhinchus larvae fed exclusively on dry diets at 2 temperatures (8.4 and 17.5°C) during a 32 d rearing experiment. The higher temperature enhanced growth significantly. Mean dry weights at the end of the experiment were 3.6 mg (SD = 1.07, range 1.4 to 5.7 mg) and 31.5 mg (SD = 21.9, range 3.6 to 96.0 mg), and mean standard lengths were 17.7 ± mm (SD = 1.6, range 11.8 to 19.6 mm) and 25.5 mm (SD = 4.2, range 17.0 to 35 mm), respectively. Significant responses to temperature were also found in the nucleic acids. However, these differences were not as remarkable and gave indications that differences in protein growth between treatments was based on protein biosynthesis being driven by the activity of the ribosomes, rather than their number. The use of the degree-day approach to normalize the data clearly showed the temperature-dependence of somatic growth. Only small differences in growth and nucleic acid content were observed in the comparable range of the first 280 degree-days in fed fish. In non-fed fish, the starving potential was very similar (approx. 350 degree-days). Shifts in growth pattern from predominantly hyperplasia to predominantly hypertrophy were detected, with ongoing growth at both temperatures.