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A feeding trial was conducted to test the effect of dietary glutamate and glycine supplementation on growth potential, nutritional utilization and digestion, and energy metabolism of rainbow trout (Oncorhynchus mykiss). Four isonitrogenous and isolipidic diets (2 × 2 factorial design) were formulated with glutamate (Glu) and glycine (Gly) as the main factors. The protein source consisted mainly of black soldier fly larvae meal (40% inclusion in all diets) for its low-level content of glutamate and glycine. Triplicate groups of rainbow trout (30 fish/tank with mean initial body weight of 87.5 g ± 0.9) were restrictively fed by hand twice a day for 6 weeks. Dietary Glu and Gly supplementation did not affect feed intake, specific growth rate, or the feed conversion ratio. Whole body protein, lipid, amino acid and fatty acid composition were also unaffected by the supplementation of Glu and Gly in the diets. Dietary Gly supplementation increased the apparent digestibility of amino acids. Furthermore, supplementation with Gly, increased the concentration of serine, glycine, tryptophane, tyrosine and citrulline, while supplementation with Glu in the diets increased the concentration of hydroxy-proline and β-alanine in the serum of rainbow trout. Both dietary Glu and Gly supplementation improved the digestibility of the fatty acids. The expression of genes involved in the hepatic bile acid synthesis (e.g., apical sodium dependent bile acid transporter and organic solute transporter) were affected by supplementation of Gly in the diets. In general, this study showed that dietary supplementation with Gly and Glu improved the nutrient digestibility in rainbow trout.

The establishment of the early-life gut microbiota plays an important role in fish development and influences the host's health status and growth performance. Different rearing conditions can impact the initial colonization of the gut microbiota, while the addition of probiotics may also affect such colonization. However, how this may affect fish larvae survival and growth remains largely unexplored. In this study, 3-day old Nile tilapia embryos were hatched until 9 days post fertilization (dpf) in three systems, including one flow-through system (FTS) and two identical recirculating aquaculture systems (RAS). When feeding started at 10 dpf, tilapia larvae in the FTS and one of the RAS were fed with a control diet, while larvae in the second RAS were fed with the control diet coated with B. subtilis spores (RASB). The feeding trial lasted 26 days, from larvae to fry stage, during which the survival, growth performance and gut microbiota were analyzed. The larvae reared in FTS showed significantly lower survival than those in RAS and RASB, while no differences were observed in fish growth and apparent feed conversion ratio between treatments. Different rearing systems resulted in different gut microbiota compositions, which strongly correlated with the survival rate and standard body length at harvest. Cetobacterium was enriched in RAS and RASB, while was barely detected in the gut of FTS-reared tilapia fry. Probiotic supplementation increased the relative abundance of beneficial Bacillus in fish gut. Our findings indicate that rearing fish larvae in RAS supports better survival compared to FTS, while dietary probiotic supplementation further modulates the gut bacterial composition and stimulates presence of beneficial bacteria during early life.

Background Fish gut microbial colonisation starts during larval stage and plays an important role in host’s growth and health. To what extent first colonisation could influence the gut microbiome succession and growth in later life remains unknown. In this study, Nile tilapia embryos were incubated in two different environments, a flow-through system (FTS) and a biofloc system (BFS); hatched larvae were subsequently cultured in the systems for 14 days of feeding (dof). Fish were then transferred to one common recirculating aquaculture system (RAS1, common garden, 15–62 dof), followed by a growth trial in another RAS (RAS2, growth trial, 63–105 dof). In RAS2, fish were fed with two types of diet, differing in non-starch polysaccharide content. Our aim was to test the effect of rearing environment on the gut microbiome development, nutrient digestibility and growth performance of Nile tilapia during post-larvae stages. Results Larvae cultured in the BFS showed better growth and different gut microbiome, compared to FTS. After the common garden, the gut microbiome still showed differences in species composition, while body weight was similar. Long-term effects of early life rearing history on fish gut microbiome composition, nutrient digestibility, nitrogen and energy balances were not observed. Still, BFS-reared fish had more gut microbial interactions than FTS-reared fish. A temporal effect was observed in gut microbiome succession during fish development, although a distinct number of core microbiome remained present throughout the experimental period. Conclusion Our results indicated that the legacy effect of first microbial colonisation of the fish gut gradually disappeared during host development, with no differences in gut microbiome composition and growth performance observed in later life after culture in a common environment. However, early life exposure of larvae to biofloc consistently increased the microbial interactions in the gut of juvenile Nile tilapia and might possibly benefit gut health.

Effects of the type of dietary non-protein energy source on the size and composition of the total body bile acid pool, on faecal bile acid loss and on bile acid synthesis were investigated in rainbow trout. Two diets were formulated (similar DP:DE ratio) that differed in the inclusion of either maize starch (Starch) or rapeseed oil (Fat) as main non-protein source. Fish were fed to satiation for 44 days. Type of non-protein energy source did not substantially affect the body bile acid pool composition. However, feeding the Starch diet resulted in a larger total body bile acid pool size compared with the Fat diet, and this despite enhanced faecal bile acid loss when feeding the Starch diet that was related to more faeces being produced. Bile acid synthesis in fish fed the Starch diet was more than two times higher compared with fish fed the Fat diet. The difference in body bile acid pool size between diets suggests upregulation of bile acid synthesis in fish fed the Starch diet beyond the level needed to compensate for the higher faecal bile acid loss and/or downregulation of bile acid synthesis in fish fed the Fat diet. The underlying mechanisms for this difference in synthesis need further investigation.

The objective of this study was to investigate the effect of the gas: liquid ratio (GLR) on carbon dioxide removal and heat loss across a forced-ventilated trickling filter (TF) used as bioreactor. The effect of 8 different GLRs (1.2–15.7) on carbon dioxide removal and heat loss from water passing a trickling filter were tested in random order. The TF was part of a pilot scale fresh water intensive recirculating aquaculture system and had a media bed height of 1.8 m, a diameter of 0.9 m, a specific surface area of 200 m2/m3 and was operated at a fixed hydraulic surface load of 9.42 m3/m2.h, a mean inlet water temperature of 27.0–27.5 °C, and a mean inlet pH ranging from 7.02–7.12. For each GLR, CO2 removal from water was calculated from CO2 measurements in the inlet and outlet air of the TF using an online infrared probe (developed for agriculture and greenhouses) and from 12 water measurements done once every 3 h during 2 subsequent days. The water inlet CO2 concentrations varied between 4.5–9.0 mg/L (calculated from total inorganic carbon and pH). The results showed that the CO2 removal from water to the air in a trickling filter increased from 3.1 mg/L at a GLR of 1 to 6.1 mg/L for a GLR of 15. The relationship between GLR and CO2 removal could be described as an asymptotic curve with the following equation C O 2 r e m o v a l f r o m w a t e r ( m g L ) = 6.44 - 5.82 ( 0.648 G L R ) ; (R2 = 0.91). A linear relation between total heat loss across the TF and the applied GLRs showed an average total heat loss of 33.2 kJ per cubic meter of air for all GLRs. Of the total heat loss on average 91 % was latent heat loss and 9 % sensible heat loss. This study showed that: (1) GLRs above 5 will not result in significant additional CO2 removal; (2) energy loss at a GLR of 5 is, under our experimental conditions with a mean inlet air temperature of 22.6 °C and an mean inlet air relative humidity of 66.9 %, 26.0 kJ per g CO2 removed.

There is a recognized need for mineralizing aquaculture-derived sludge in aquaponics systems in order to reduce waste production. Many recent studies of aquacultural waste treatment have focused only the production of biogas as opposed to the potential for mineralization of nutrient-rich sludge. Upflow anaerobic sludge blanket (UASB) reactors provide one possible solution for breaking down sludge into bioavailable nutrients that can subsequently be delivered to plants. As such, this study examines the mineralization performance of sequential UASB reactors that are designed with an expanded granular sludge bed (EGSB) and compared to standard aerobic and anaerobic batch reactors. Results of our experiments demonstrate that only chemical oxygen demand reduction is significantly different. An unexpected drop in pH of one of the three reactor systems revealed that a pH below 6 was able to significantly increase the mineralization and mobilization of nutrients. Approximately 25% of phosphorus, potassium, and calcium could also be recovered from the sludge under lower pH conditions, as compared to the mineralization performance of standard UASB reactors running at a higher pH. However, the opposite effect was observed with respect to organic sludge reduction, where diminished performance was observed in the low-pH reactor. The current study implies that anaerobic reactors operating at low pH can potentially contribute towards improved nutrient recovery in multi-loop aquaponics systems and reduction of additive agents for pH control of the hydroponic subsystem.

To assess the potential for improving sustainability and efficiency in an important Vietnamese finfish culture, we quantified sustainability indicators for 2 downstream and 2 upstream 3 to 4 m deep ponds for striped catfish Pangasianodon hypophthalmus (Sauvage, 1878) production along the Mekong River, Vietnam. The resources used per kg fish produced were (downstream vs. upstream): 2.8 vs. 7.1 m3 water, 0.04 vs. 0.14 kWh energy, 0.06 vs. 0.11 h human labour and 0.06 vs. 0.15 g antibiotics (p < 0.05). The feed conversion ratio (kg feed dry matter per kg fish wet weight) was 1.50 vs. 1.65 and feed dry matter utilization efficiency was similar (28 vs. 30%) for both types of ponds (p > 0.05). Nitrogen and phosphorus utilization efficiencies were 44 vs. 40% for nitrogen (p < 0.05) and 17.7 vs. 17.6% for phosphorus (p > 0.05). An advantage of production in deep ponds is the breakdown of organic matter through denitrification and fermentation. Based on the difference between feed input and monitored outputs and sinks, an estimated 29–37% of dry matter volatilized as CO2 and 30–34% of nitrogen was lost as N2. A major disadvantage of the deep pond production system is its dependency on water exchange with the Mekong River, resulting in low biosecurity, which is aggravated by the lack of availability of disease-free fingerlings. Dealing with disease caused farmers to apply suboptimal feeding rations and to lengthen the culture period. Developing (semi-)closed systems that adopt elements from recirculation technology could improve both culture performance and sustainability.

The aquatic metabolic unit used in this study was cofounded by The Netherlands Organization for Scientific Research (code 805-34.025). Currently it is part of a large-scale EU research facility program. The study was funded by the EU-FP7 project AQUAEXCEL (262336). LM is currently supported by a Fundação para a Ciência e a Tecnologia (FCT) research position (IF/01314/2014/CP1231/CT0001) from Portugal. This study had also the support of FCT through strategic projects UID/Multi/04423/2019 and UID/MAR/04292/2013 granted to CIIMAR and MARE, respectively. SN has support from FCT grant (SFRH/BPD/94500/2013). This work was also supported by the Integrated Programme of SR&TD “Smart Valorization of Endogenous Marine Biological Resources Under a Changing Climate” (reference Centro01-0145-FEDER-000018), co-funded by Centro 2020 program, Portugal 2020, European Union, through the European Regional Development Fund. In aquaculture, fish may be exposed to sub-optimal rearing conditions, which generate a stress response if full adaptation is not displayed. However, our current knowledge of several coexisting factors that may give rise to a stress response is limited, in particular when both chronic and acute stressors are involved. This study investigated changes in metabolic parameters, oxidative stress and innate immune markers in a rainbow trout (Oncorhynchus mykiss) isogenic line exposed to a combination of dietary (electrolyteimbalanced diet, DEB 700 mEq Kg−1 ) and environmental (hypoxia, 4.5 mg O2 L −1 ) challenges and their respective controls (electrolyte-balanced diet, DEB 200 mEq Kg−1 and normoxia, 7.9 or mg O2 L −1 ) for 49 days. At the end of this period, fish were sampled or subjected to an acute stressor (2 min of handling/confinement) and then sampled. Feeding trout an electrolyte-imbalanced diet produced a reduction in blood pH, as well as increases in cortisol levels, hepato-somatic index (HSI) and total energy content in the liver. The ratio between the lactate dehydrogenase (LDH) and isocitrate dehydrogenase (IDH) activities decreased in the liver of trout fed the DEB 700 diet, but increased in the heart, suggesting a different modulation of metabolic capacity by the dietary challenge. Several markers of oxidative stress in the liver of trout, mainly related to the glutathione antioxidant system, were altered when fed the electrolyteimbalanced diet. The dietary challenge was also associated with a decrease in the alternative complement pathway activity (ACH50) in plasma, suggesting an impaired innate immune status in that group. Trout subjected to the acute stressor displayed reduced blood pH values, higher plasma cortisol levels as well as increased levels of metabolic markers associated with oxidative stress in the liver. An interaction between diet and acute stressor was detected for oxidative stress markers in the liver of trout, showing that the chronic electrolyte-imbalance impairs the response of rainbow trout to handling/confinement. However, trout reared under chronic hypoxia only displayed changes in parameters related to energy use in both liver and heart. Taken together, these results suggest that trout displays an adaptative response to chronic hypoxia. Conversely, the dietary challenge profoundly affected fish homeostasis, resulting in an impaired physiological response leading to stress, which then placed constraints on a subsequent acute challenge. info:eu-repo/semantics/publishedVersion

This paper compares the quantity and quality of solid waste obtained from traditional ponds and recirculating aquaculture systems (RAS) for striped catfish and evaluates methane and compost production from these wastes. Striped catfish sludge was collected from four commercial ponds along the Mekong river and from three indoor RAS. The amount of sludge dry matter produced per kilogram of fish in ponds was 6 times higher than that in RAS. However, the concentration of nutrients in solid waste from RAS was much higher, with better compost quality and higher methane yield than that of sludge from ponds. Out of the collected 381 L biogas, the methane yield of striped catfish’s solid waste in RAS systems was 201 L per kg chemical oxygen demand (COD). In ponds, the collected 267 L biogas yielded 125 L CH4 per kg COD. The higher methane production from RAS sludge concurred with higher digestibility of COD: 58% for RAS versus 38% for ponds. The quality and quantity of methane from striped catfish sludge were lower than that for other animal manures, and the resulting electricity yield was low. Considering the higher nutrient concentration in RAS-sludge, we recommend the combination RAS and composting in reusing sludge, which is presently the best option for a more sustainable and cleaner striped catfish production system.

Oxygen limitation and dietary imbalances are key aspects influencing feed intake (FI) and growth performance in cultured fish. This study investigated the combined effects of hypoxia and dietary electrolyte balance on the growth performance, body composition and nutrient utilization in a rainbow trout (Oncorhynchus mykiss) isogenic line. Fish were fed ad libitum two experimental diets: electrolyte-balanced or -imbalanced diets (DEB 200 or 700 mEq kg−1, respectively) and exposed to normoxia or hypoxia (7.9 or 4.5 mg O2 l−1, respectively) for 42 days. DEB did not affect FI, growth performance or body composition. Nevertheless, hypoxia had a negative impact, reducing FI (6%), growth rate (8%), oxygen consumption (19%), energy (5%) and lipid (42%) contents. Digestible energy intake and heat production were higher in normoxic fish (40% and 23%, respectively), retaining 64% more energy in lipid or protein. Hypoxia reduced the apparent digestibility of dry matter, ash, protein, lipid, carbohydrates and energy. Trout fed DEB 700 diet were energetically less efficient, reflected in higher heat production and energy requirements for maintenance. FI was inhibited by low dissolved oxygen levels, but not by electrolyte-imbalanced diet, in spite of the higher energy requirements for maintenance. This study highlights the importance that dietary-electrolyte content and DO levels have on energy balance and growth performance when fish are fed to satiation.

We evaluated the effect of low pH and low and high total ammonia nitrogen (TAN) concentrations on the physiology, stress status and the growth performance of turbot in RAS. Two experiments were conducted. In Experiment 1, turbot (466 g) were grown at control (pH 7.5; TAN ~0.5 mg/L) or low pH and high TAN (pH 5.7; TAN ~50 mg/L) for 55 days. In Experiment 2, turbot (376 g) were grown at control (pH 7.5; TAN ~0.5 mg/L), low pH and low TAN (pH 5.7; TAN ~5 mg/L) or low pH and high TAN (pH 5.7; TAN ~50 mg/L) for 59 days. In Experiment 1, final body weight, feed intake and growth were significantly lower and FCR significantly higher in turbot exposed to low pH and high TAN. In Experiment 2, only growth was significantly lower in turbot exposed to treatment low pH and high TAN as compared to fish in the control treatment and low pH and low TAN. Osmoregulation and stress indicators measured were within normal levels. In conclusion, turbot grew equally well in a water pH of 7.5 or 5.7 provided a low TAN. In contrast, low pH combined with a high TAN impaired turbot performance. The effect of low pH on physiology, stress status and growth performance of turbot (Psetta maxima L.) cultured in recirculating aquaculture systems

The effect of a high and low non-starch polysaccharide diet (HNSP and LNSP diet) and ultrasound treatment on particle size distribution and carbon bioavailability in fecal waste of rainbow trout (Oncorynchus mykiss) was studied. Feces were collected from four flow-through fish tanks, two tanks fed the HNSP diet and two the LNSP diet. The collected feces were sonicated (disintegrated) in duplicate with high-intensity (0.6 W/ml), low-frequency (f = 20 Hz) ultrasound at five different energy levels (0.6 W/ml for 0, 0.25, 1, 4, and 16 min). The particle size distribution of the treated feces samples was measured by wet sieving (1000, 500, 200, 100, 63, 36, 1.2 μm screen size) and total suspended solids (TSS) measurement. Carbon bioavailability in sonicated fecal waste samples was determined with oxygen uptake rate (OUR) tests. The results showed that: (1) feces from the HNSP diet contained significant more particulate material and bigger particles; (2) carbon bioavailability was almost three times higher in untreated LNSP feces when compared with HNSP feces; (3) almost 50% of HNSP feces could have been recovered on a microscreen of 36 μm after wet sieving, whereas it was only 10% for LNSP feces; (4) the production of small particles (1.2–36 μm), which could pass a drum filter screen and potentially accumulate in RAS, was approximately 50 g/kg feed, showing no significant differences between diets; (5) sonication increased fecal dry matter below 36 μm (p = 0.015), but it had no significant effect on the median particle size; (6) sonication increased carbon bioavailability with 7–10% for the HNSP feces (p = 0.037); (7) fecal particles withstood up to 16 min sonication at an intensity of 0.6 W/ml and a frequency of 20 Hz corresponding to specific energy input of 20,000 kJ/kg DM without major changes in particle size distribution. The results of this study indicate that the applied ultrasound treatment of fecal waste is not an effective method to increase short-term carbon bioavailability.

The accumulation of steroids released by fish in recirculating aquaculture systems (RAS) may potentially influence their physiology and behavior. The present study examined the release rate of cortisol and testosterone by Nile tilapia, Oreochromis niloticus, and their accumulation in six identical lab scale RAS operated at different water exchange rates (150 L/kg feed/day, (LowRAS) and 1500 L/kg feed/day, (HighRAS)) and how steroid accumulation is affected by grading and weighing induced stress. Water cortisol and testosterone concentrations during the experimental period ranged between 1.0 and 5.1 ng/L and between 1.4 and 9.4 ng/L, respectively. Water cortisol concentration was 34% and 43% significantly higher in LowRAS when compared with HighRAS at week 3 and week 4, whereas for water testosterone concentration the two treatments did not differ. Overall steroid release rates were 0.02 +/- 0.02 and 0.04 +/- 0.02 ng/g body weight/h for cortisol and testosterone, respectively. Two hours after grading and weighing water cortisol concentration increased 30% in the LowRAS whereas water testosterone concentration remained unchanged. Calculated cortisol and testosterone discharge from the system were, respectively, 87% and 89% lower in LowRAS than in HighRAS. These findings show that reduced water usage and acute stressors can induce significant accumulation of cortisol in the rearing water at levels close to olfactory detection. Accumulation of metabolites, in particular steroids, should be taken in consideration when designing and managing RAS to prevent exceeding allowable concentrations. Statement of relevance: RAS containing Nile tilapia, cortisol and testosterone are released to the culture water. Water cortisol concentration depends on the water exchange rate that is used and higher concentrations are expected in lower water exchange rates. Fish grading and weighing has a short-term effect on the water cortisol concentrations, after which concentrations return to the basal levels. These findings show that reduction in water usage and fish handling may lead to a significant increase of hormones in the rearing water, which emphasizes the importance of accounting for steroids in the design and management of RAS. (C) 2016 Elsevier B.V. All rights reserved. Portuguese institution FCT - Fundacao para a Ciencia e Tecnologia (QREN-POPH - Advanced Training - European Social Fund) [SFRH/BD/65673/2009] Portuguese institution FCT - Fundacao para a Ciencia e Tecnologia (QREN-POPH - Advanced Training - national funds MEC) [SFRH/BD/65673/2009]

Steroids accumulate in recirculating aquaculture system (RAS), although explanatory factors for such accumulation are still poorly explored. This study investigated the effect of water exchange rate and pH in six replicated RAS on the concentration of the stress hormone cortisol in rainbow trout blood plasma and in the holding water and of the sex steroids testosterone, 11-ketotestosterone (11-KT) and 17,20-dihydroxypregn-4-en-3-one (17,20-P) over a 70-day experimental period. Three combinations of water exchange rate and pH were used each treatment, with two replications: (i) high water exchange rate (+/- 1700Lkg(-1) feed) and neutral pH (+/- 7.3), (ii) low water exchange rate (+/- 500Lkg(-1) feed) and neutral pH (+/- 7.3) and (iii) low water exchange rate (+/- 500Lkg(-1) feed) and low pH (+/- 5.8). Plasma cortisol concentrations at day 70 were higher (24.4 +/- 9.5ngmL(-1)) for fish kept at low pH when compared to fish kept at neutral pH (12.0 +/- 0.1 and 8.7 +/- 0.2ngmL(-1)). Water cortisol and testosterone concentrations at day 35 were higher at low pH than at neutral pH, whereas water 11-KT and 17,20-P did not differ among treatments. At day 70, there were no significant differences between low and high pH. These results demonstrate that low pH contributes to increased plasma cortisol concentrations and to its accumulation in water, possibly indicating a stress response to low pH. The higher concentration of testosterone but not of the other sex hormones point to unspecified reproductive effects that need further investigation. PhD scholarship - FCT - the National Foundation for Science and Technology of Portugal through QREN-POPH Advanced Training European Social Fund and National funds (MEC) [SFRH/BD/65673/2009] info:eu-repo/semantics/publishedVersion

Denitrification on internal carbon sources offers the advantage to control nitrate levels in recirculating aquaculture systems (RAS) by using the fecal carbon produced within the husbandry system. However, it is not clear to which extent fecal carbon can be utilized by the microbial community within a denitrification reactor. Especially fibers can hamper the bioavailability of carbon in fecal waste. Therefore, this study investigated the nitrogen removal capacity of a denitrification reactor using fecal waste with a high fiber content as the only carbon source in RAS. Furthermore, we investigated to which extent fibers were utilized as a carbon source within the reactor. Four identical small-scale RAS (V = 460 L) were stocked with 25 rainbow trout of ~ 110 g, and operated at a water exchange rate of ~ 200 L/kg of feed DM. Two RAS served as controls without denitrification and two RAS were upgraded with an upflow sludge blanket denitrification reactor (V = 10.5 L). During the six weeks of experiment, we determined COD (chemical oxygen demand, measure for organic carbon) and N balances for all systems and analyzed the composition of the collected solids. The denitrification reactors were able to remove 19 g N/kg of feed DM, or 48% of the metabolic nitrogen waste produced by the fish. Based on the COD balances, 44% of the supplied fecal COD was degraded in the reactor. Hemicellulose and cellulose degradability was ~ 50%, accounting for 45% to the total degraded COD. Under steady state conditions, 4.4 g of biodegradable COD needed to be oxidized to reduce 1 g of nitrogen, indicating respiratory COD losses of approximately 50%. This experiment successfully demonstrated that denitrification on internal carbon sources using a high fiber diet could remove half of the nitrogen waste produced by the fish. Although fibers limited carbon bioavailability, half of the cellulose and hemicellulose present in the fecal waste was utilized in the denitrification reactor.

The accumulation of substances in Recirculating Aquaculture Systems (RAS) may impair the growth and welfare of fish. To test the severity of contaminants accumulated in RAS, early-life stages of fish were used. Ultrafiltered water from two Recirculating Aquaculture Systems (RAS), one RAS with a high accumulation of substances (water exchange rate 30 L/kg feed/day) and one RAS with a low accumulation of substances (water exchange rate 1500 L/kg feed/day), was used to incubate eggs and rear larvae of common carp Cyprinus carpio . A broad range of read-out parameters was used to determine the effect of accumulation level on the development of the early-life stages; from hatching dynamics to larvae length and dry weight. The water quality (temperature, pH, dissolved O 2 , conductivity, total bicarbonate, ortho-phosphate-P, TAN, NO 2 − –N, NO 3 − –N and minerals) was compared between the 2 treatments. Carp eggs developing in the high-accumulation water had higher mortality percentages (both for eggs and larvae), reduced hatching percentages, delayed hatching dynamics and reduced larvae length and body weight. However, these larvae exhibited fewer deformities than larvae incubated in the low-accumulation water. Furthermore, an accelerated development both of the embryo (appearance of heart beat, pectoral fin bud and tail movement) and yolk-sac larvae (depletion of the yolk sac) was observed in the high-accumulation water. The high-accumulation water had significantly lower pH and higher conductivity, NO 2 − –N, NO 3 − –N and ortho-phosphate-P. Most of the minerals (As, Cu, Mn, Ni, Zn, K, Mg, Na, P and S) including heavy metals, were present at a higher concentration in the high-accumulation water. The influence of these parameters on the embryonic and larval development of fish is discussed. It is suggested that in the high-accumulation water, the concentration of ortho-phosphate-P, nitrate and of the heavy metals arsenic and copper is likely to have impaired the embryonic and larval development and therefore deserves further research as potential growth inhibiting factors in RAS.

It has been suggested that fish cultured in recirculating aquaculture systems (RAS) grow less as compared with fish cultured in flow-through systems due to the accumulation of substances. In the Netherlands, the commercial culture of Nile tilapia Oreochromis niloticus in 300 and 600 MT's systems is done exclusively in RAS operated at water exchange rates as low as 30 L/kg feed/day due to nitrate control by single-sludge denitrification reactors. The use of such nearly closed RAS raises the question whether growth retardation (GR) is present in Nile tilapia. This study is the first to investigate the existence of growth retardation in Nile tilapia by comparing the growth, feeding behaviour and stress response of Nile tilapia cultured in RAS with different levels of substances accumulated. Three RAS, operated at 30 L/kg feed/day (HIGH accumulation), 70 L/kg feed/day (MIDDLE accumulation) and 1500 L/kg feed/day (LOW accumulation) were used. Each RAS contained 24 glass aquaria with individually housed fish. To determine whether GR is size-dependent, per RAS 3 fish size categories were tested in the 57 day experimental period: large (288.7 ± 34.2 g; N = 8), medium (162.4 ± 23.4 g; N = 8) and small (81.4 ± 21.0 g; N = 8). Experimental fish were fed ad libitum , twice per day. Feeding behaviour was determined once per week and was measured as the time taken by each fish to eat the first pellet (latency, LAT) and the total time spent feeding (total feeding time, TFT). Temperature, pH, conductivity, alkalinity, dissolved oxygen, dissolved CO 2 , nitrogen compounds (TAN, NO 2 -N and NO 3 -N), chemical oxygen demand and orthophosphate-P were measured over time. At day 57 fish were weighed, blood sampled and returned to their tanks for an extra experimental period of 15 days, and subjected at day 72 to an acute stress followed by blood sampling. Blood was analysed for glucose and cortisol. Results showed that the water quality parameters measured in the 3 RAS (with the exception of alkalinity) were still within the optimum range for growth of Nile tilapia. Large individuals showed a tendency to grow more in the LOW treatment (2.66 ± 1.35 g/kg 0.8 /day) as compared with the MIDDLE treatment (0.93 ± 1.63 g/kg 0.8 /day). On the contrary, small individuals grew significantly less in the LOW treatment (3.60 ± 1.74 g/kg 0.8 /day) as compared with the HIGH (7.22 ± 1.58 g/kg 0.8 /day) and MIDDLE (6.82 ± 4.54 g/kg 0.8 /day) treatments. Small fish were more motivated to eat (lower latency) in the MIDDLE (4.63 ± 5.24 min) as compared with the LOW treatment (8.94 ± 6.41 min). In the HIGH accumulation treatment higher glucose levels were observed in the small fish, before and after acute stress, as compared with the LOW accumulation treatment. In conclusion, this study showed that the extent to which the accumulation of substances in RAS affects growth depends on fish size: large individuals show a trend towards growth retardation in the highest accumulation RAS while small individuals, on the contrary, seem to grow better in such systems.

The kinetics and design of a suspended bacteria growth reactor, which can be integrated in a 100MT African catfish farm, were determined. Such a reactor converted nitrogen (N) and phosphorus (P) from RAS effluents into heterotrophic bacteria biomass. The determined kinetics were: yield = 0.537 g VSS/g C; endogenous decay coefficient = 0.033 h −1 ; maximum specific growth rate = 0.217 h −1 ; half-velocity constant = 0.025 g/l; maximum rate of substrate utilization = 0.404g C/g VSS h. A reactor integrated in a 100MT farming facility would have a volume of 11 m 3 , based on a minimum HRT of 6 h. The kinetics and reactor design were integrated in a model to predict the VSS production (volatile suspended solids as measure of bacteria biomass) and nutrient conversions. The VSS production was on average 187 ± 2 g VSS/kg feed and the inorganic nutrients (N and P) were removed with an efficiency of 85 ± 3.0% and 95 ± 2.5%, respectively. A carbon (C) supplementation level of 455 g C/kg feed was required to ensure optimal C:N ratios for heterotrophic bacteria production. The production of heterotrophic bacteria biomass is, therefore, a prospective tool to lower nutrient discharge and to increase nutrient retention and sustainability of RAS in the future.

One of the challenges that Recirculating Aquaculture Systems (RAS) are still facing is the risk that in RAS fish grow less than in flow-through systems due to the accumulation of substances originating from feed, fish or bacteria associated with the water re-use. The present study investigated whether RAS with high and low accumulation levels of these substances affect feed intake and growth of Nile tilapia Oreochromis niloticus, African catfish Clarias gariepinus, and European eel Anguilla Anguilla. One-hundred and twenty individuals of each species were used (start body weights: Nile tilapia 264.8 ± 8.3 g; African catfish 253.2 ± 2.1 g and European eel 66.6 ± 1.3 g). For a period of 39 days, growth and feed intake were compared between high and low accumulation RAS. HIGH accumulation RAS was designed for maximal accumulation of substances in the water by operating the system at nearly-closed conditions (30 L/kg feed/d), using mature biofilters and high feed loads; and (2) LOW accumulation RAS was designed to be a proxy for flow-through systems by operating at high water exchange rates (1500 L/kg feed/d), new biofilters and low feed load. HIGH accumulation RAS induced a reduction in feed intake (42%) and growth (83%) of Nile tilapia, as compared to systems that are a proxy for flow-through conditions. This effect was not observed in European eel and African catfish. The cause of this reduced feed intake and growth rate of Nile tilapia is still unclear and should be addressed in further studies.

This paper analyses nutrient conversions, which are taking place in integrated intensive aquaculture systems. In these systems fish is cultured next to other organisms, which are converting otherwise discharged nutrients into valuable products. These conversions are analyzed based on nitrogen and phosphorous balances using a mass balance approach. The analytical concept of this review comprises a hypothetical system design with five modules: (1) the conversion of feed nutrients into fish biomass, the “Fish-Biomass-Converter”; (2) the separation of solid and dissolved fish waste/nutrients; the “Fish-Waste-Processor”; (3) the conversion of dissolved fish waste/nutrients, the “Phototrophic-herbivore-Converter”; (4 and 5) the conversion of solid fish waste, the “Bacterial-Waste-Converter”, or the “Detrivorous-Converter”. In the reviewed examples, fish culture alone retains 20–50% feed nitrogen (N) and 15–65% feed phosphorous (P). The combination of fish culture with phototrophic conversion increases nutrient retention of feed N by 15–50% and feed P by up to 53%. If in addition herbivore consumption is included, nutrient retention decreases by 60–85% feed N and 50–90% feed P. This is according to the general observation of nutrient losses from one trophic level to the next. The conversion of nutrients into bacteria and detrivorous worm biomass contributes only in smaller margins (e.g. 7% feed N and 6% feed P and 0.06% feed N 0.03 × 10−3% feed P, respectively). All integrated modules have their specific limitations, which are related to uptake kinetics, nutrient preference, unwanted conversion processes and abiotic factors.

The need for unbiased digestibility estimation has led to the development of a number of faeces collection methods. However, there is still a large variation in apparent digestibility coefficient (ADC) of diet/ingredients between these methods. This study investigated the impact of dietary quality on ADC and faeces recovery measurements obtained by two faecal collection methods (Choubert and settling tank). The fish were fed five diets: a control diet as high-quality diet, two levels of dried duckweed (20% and 40%) and two levels of fresh duckweed (20% and 40%), which were added to the control diet, as low-quality diets. Apparent digestibility coefficient estimates were highly correlated (r>0.95) between both faecal collection methods. For all diets the ADC estimates were higher when using settling tanks compared with Choubert collectors. For the control diet differences in ADC between two collectors were small, but by inclusion of duckweed these differences increased. The recovery percentage of faeces was not correlated between both faecal collection methods (r=0.22, P=0.41). The estimated recovery percentage of faeces by settling tanks was about three times higher than by Choubert. In conclusion, the differences in ADC and faeces recovery measured by two faecal collection methods depend on diet quality

Abstract Gonadal development and spawning behaviour of artificially-matured European eel (Anguilla anguilla L.) was studied. Treatment of males with Human Chorionic Gonadotropin (HCG; 1 IU g/week) resulted in a Gonado-Somatic Index (GSI) of 10.88 ± 3.39 and spermiation. Treatment of females with carp Pituitary suspension (cPs) (20 mg cPs/kg body weight per week) resulted in oogenesis with a GSI of 20.0 ± 11.3 (n = 7), and the number of eggs per female was 1874 * 103 ± 1116 * 103; (n = 7). Ovulation of the females was induced with 17α, 20β dihydroxyprogesteron (DHP) at 2 μg/g bodyweight. Eggs of European eel were found to be non-sticky and typically pelagic. Maximum speed of eggs rising to the surface in a water column was 2.24 ± 0.33 metres (m) per hour (h). To study behaviour in a qualitative way, two females were used together with three groups of three males. During a 283 minute (min) observation of the two females, we observed female-female interaction: 'lethargic behaviour' (33.6%) vs. 'cruising together' (66.4%). In the period when males and females were together (188 min), we observed 'approaching the head region of the female' (57.7%), 'touching the operculum' (39.4%), or 'approaching the urogenital area' (2.9%) by the males (total 725 seconds (s)). Sperm release in the presence of a female took 115 s of the total approaching time of 725 s (15.9%), while in the case of male-male interaction this was only 15 s of the total period of 116 s (12.9%). Induced spawning behaviour of eels was collective and simultaneous, corresponding to spawning in a group. This is the first time group spawning behaviour has ever been observed and recorded in eels.

Little information is available on steroid concentrations in the rearing water of aquaculture systemsand whether they accumulate in recirculating aquaculture systems (RAS). Therefore this study aimedat determining (1) the concentrations and variation of cortisol and sex steroids in RAS, (2) the con-tribution of fish rearing conditions to steroid concentrations in seven commercial RAS. Each RAS wassampled twice at three different points: (1) make-up water; (2) influent and (3) effluent of the rear-ing unit. The results showed significant higher steroid concentrations in the influent and effluent whencompared with the make-up water. On average cortisol concentration was 15.7% higher in the efflu-ent when compared with the influent. Mean steroid concentrations in the rearing unit effluent variedbetween: 3.8–217.0 ng/L for cortisol, 3–12.5 ng/L for testosterone, 0.9–7.1 ng/L for 11-ketoteststerone and1.8–12.8 ng/L for 17,20 -dihydroxypregn-4-en-3-one. Stocking density, Total Ammonia-Nitrogen con-centration and orthophosphate-P concentration (a measure of make-up water usage) showed a positivecorrelation with sex steroids in the water. The steroid concentrations from the present study were ordersof magnitude lower than initial estimations indicating a water treatment efficiency of >99%. The resultssuggest that an intensification of fish production through decrease of make-up water use and increaseof stocking density will lead to a build-up of steroids in the water. Although intensification is critical forthe economical success of RAS, this ultimately could affect fish performance as steroids accumulates inthe water of RAS at levels that can potentially be detected by some fish species. Institution FCT – Fundação para a Ciência e Tecnologia

This study investigated the effect of dietary carbohydrate composition on the production, recovery and degradability of fecal waste from rainbow trout (Oncorhynchus mykiss) in recirculating aquaculture systems (RAS). Dietary carbohydrate composition was altered by substituting starch with non-starch polysaccharides (NSP) while keeping the diets isonitrogenous and isoenergetic. We tested a high starch, low NSP (LNSP) and a low starch, high NSP (HNSP) diet in six identical, small-scale RAS (V = 460 L). Each diet was tested in three independent systems over a period of six weeks. Shifting dietary carbohydrates from starch to NSPs resulted in a 50% increase in the production of chemical oxygen demand (COD) based on digestibility. Fecal waste recovery showed a 40% increase in HNSP treatments when compared with LNSP. Consequently, the COD output from HNSP systems doubled from 91 g to 194 g of COD per kg feed when compared with LNSP. Although COD production was higher in HNSP systems, the COD load on the biofilters was significantly lower when compared with LNSP systems. COD-to-nitrogen (COD/N) ratios in the biofilter load were 1.7 ± 0.2 and 2.2 ± 0.2 g COD/g N for HNSP and LNSP, respectively. Shifting the dietary carbohydrate composition from starch to NSPs decreased the biodegradability of fecal COD from 66.3% to 43.7% (P

Migrating European eel ( Anguilla anguilla L.) glass eels were collected annually in the Garonne Basin (France) during 1987–1989 and subsequently raised under freshwater conditions. In 1990 gonadal development was determined and after about 1 year it was re-examined to check if sex ratios had changed. Undifferentiated gonads were seldom found, and were mainly present in the youngest year sample 89-1 + (numbers refer to year of catch followed by freshwater age). This gonadal stage can develop directly into an ovary. Differentiation of male sex proceeded via an intersexual stage with male and female sex cells. Phenotypic expression of female gonadal sex was considered as being gonochoristic and differentiated. The female gonad gradually developed towards a fourth stage ovary, characterised by the presence of follicles containing oocytes arrested in the late prophase of meiosis. Yolk formation in oocytes was not observed. The intersexual progression to males was termed pre-maturational sex change. The testes of full grown males contained numerous tubules with spermatogonia in mitotic arrest. The percentage of intersexual eels in 1990 of the youngest year sample (89-1+) was very high (91%). Most of these animals differentiated into males in 1991 (89-2+). The percentages of undifferentiated and intersexual eels in the older year samples of 1988 and 1987 examined in 1990 were low, showing that sex differentiation in most animals had occurred after 2–3 years. The sex ratios expressed as males/(males + females) were 64% (88-2+) and 76% (88-3+); 84% (87-3+) and 83% (87-4+). Comparison of these sex ratios suggests that sex reversal did not occur. Selective mortality and migration, population density, feeding conditions and temperature are reviewed as possible factors controlling sex differentiation in European eel.

The drumfilter effluent from a recirculation aquaculture system (RAS) can be used as substrate for heterotrophic bacteria production. This biomass can be re-used as aquatic feed. RAS effluents are rich in nitrate and low in total ammonia nitrogen (TAN). This might result in 20% lower bacteria yields, because nitrate conversion into bacteria is less energy efficient than TAN conversion. In this study the influence of TAN concentrations (1, 12, 98, 193, 257 mg TAN/l) and stable nitrate-N concentrations (174 ± 29 mg/l) on bacteria yields and nitrogen conversions was investigated in a RAS under practical conditions. The effluent slurry was supplemented with 1.7 g C/l sodium acetate, due to carbon deficiency, and was converted continuously in a suspended bacteria growth reactor (hydraulic retention time 6 h). TAN utilization did not result in significantly different observed yields than nitrate (0.24¿0.32 g VSS/g C, p = 0.763). However, TAN was preferred compared to nitrate and was converted to nearly 100%, independently of TAN concentrations. TAN and nitrate conversions rates were differing significantly for increasing TAN levels (p

This review deals with the main mechanisms and parameters affecting design and performance of trickling filters in aquaculture. Relationships between nitrification rates and easily accessible process parameters, like bulk phase concentration of TAN, O2, organic matter (COD), nitrite, temperature, HCO3−, pH and the hydraulic loading of the trickling filter, are discussed in relation to the design and operation of such filters. Trickling filter design procedures are presented and one of them, a model describing the nitrification performance of trickling filters by plug-flow characteristics, is discussed in greater detail. Finally, practical aspects in relation to filter design and operation are presented.

The effluent from the drumfilter of a recirculation aquaculture system was used as substrate to produce heterotrophic bacteria in suspended growth reactors. The effects of organic carbon supplementation (0, 3, 6, 8 g/l sodium acetate) and of hydraulic retention times (11-1h) on bacteria biomass production and nutrient conversion were investigated. Bacteria production, expressed as volatile suspended solids (VSS), was enhanced by organic carbon supplementation, resulting in a production of 55-125 g VSS/kg fish feed (0.2-0.5 g VSS/g carbon). Maximum observed crude protein production was approximately 100 g protein/kg fish feed. The metabolic maintenance costs were 0.08 Cmol/Cmol h, and the maximum growth rate was 0.25-0.5 h(-1). Ninety percent of the inorganic nitrogenous and 80% of ortho-phosphate were converted. Producing bacteria on the drumfilter effluent results in additional protein retention and lowers overall nutrient discharge from recirculation aquaculture systems.

The drumfilter effluent from a recirculation aquaculture system (RAS) can be used as substrate for heterotrophic bacteria production. These bacteria can be reused as aquatic feed. In RAS drumfilter effluents are organic carbon deficient for bacteria production. This is due to nitrogen accumulation in the system water. In the present experiment, a bacteria growth reactor (3.5l) was connected to the drumfilter (filter mesh size 60 μm) outlet of a recirculation system. To counteract carbon deficiency, different supplementation levels of molasses (organic carbon) were tested (carbon fluxes of 0.0, 3.2, 5.8, 7.8, 9.7 gC molasses/l/d; C:N ratios: 3.4, 6.4, 9.4, 13.0 and 16.5). The hydraulic retention time in the bacteria reactor was 6 h. For the maximum flux, the VSS and crude protein productions were about 168 gVSS and 95 g crude protein per kg feed. The maximum conversion of nitrate and ortho-phosphate was 24 g NO3−N and 4 gP/kg feed. This equals a conversion of 90% of the inorganic nitrogenous waste and 98% of the ortho-phosphate–P. Furthermore the maximum substrate removal rate and the Ks for molasses were determined (1.62 gC/l/h and 0.097 gC/l respectively). The maximum specific removal rate was 0.31 gC/gVSS/h and the related half saturation constant was 0.008 gC/l. The observed growth rate reached a maximum for C fluxes higher than 8 g/l/d. The present integration of heterotrophic bacteria production in RAS represents, therefore, an innovative option to reduce waste discharge by conversion. If the bacteria are reused as feed, system's ecological sustainability increases.

SUMMARYOne of the mysteries of the animal kingdom is the long-distance migration(5000–6000 km) of the European eel Anguilla anguilla L. from the coasts of Europe to its spawning grounds in the Sargasso Sea. The only evidence for the location of the spawning site of the European eel in the Sargasso Sea is the discovery by Johannes Schmidt at the beginning of the previous century of the smallest eel larvae (leptocephali) near the Sargasso Sea. For years it has been questioned whether the fasting eels have sufficient energy reserves to cover this enormous distance. We have tested Schmidt's theory by placing eels in swim tunnels in the laboratory and allowing them to make a simulated migration of 5500 km. We find that eels swim 4–6 times more efficiently than non-eel-like fish. Our findings are an important advance in this field because they remove a central objection to Schmidt's theory by showing that their energy reserves are, in principle, sufficient for the migration. Conclusive proof of the Sargasso Sea theory is likely to come from satellite tracking technology.

This study shows that alternatives for fishmeal in a fish diet affect not only fish growth but also faeces stability and nitrogen (N) and phosphorus (P) waste production. Wheat gluten diet (WGD), soybean meal extract diet (SBE), soybean meal diet (SBM), duckweed diet (DWD) and single-cell protein diet (SCP) were evaluated as a fishmeal replacement on a 15% weight weight(-1) basis in tilapia diets. Fishmeal replacement affected dry matter (dm), protein, ash and P digestibility significantly. Faeces recovery (6.8-11.2%) was not significantly affected, although the amount of non-recovered faeces and total faeces showed significant differences. Duckweed diet and SCP resulted in the largest amounts of non-recovered and total faeces (199-210, 224-225 g dm kg(-1) feed dm). Compared with fishmeal diet (FMD), the WGD and SBE resulted in similar growth, but higher non-faecal N losses (471-495 vs. 416 g N kg(-1) N). Soybean meal diet, DWD and SCP resulted in lower growth but less non-faecal loss (409-450 g N kg(-1) N). The DWD and FMD had the highest N retention (480 g N kg(-1) N) compared with the other diets (431-451 g N kg(-1) N). Carbon retention, faecal and non-faecal losses and P retention were similar for all diets (302-358, 142-176 and 489-523g C kg(-1) C, 606-704 g P kg(-1) P). Phosphorus faecal loss was lower for all diets (329-381 g P kg(-1) P) than for the FMD (401 g P kg(-1) P).

Migrating European eel (Anguilla anguilla L.) glass eels were collected annually in the Garonne Basin (France) during 1987–1989 and subsequently raised under freshwater conditions. In 1990 and 1991 sex differentiation, gonadal development, length/weight and eye size were studied in relation to metamorphosis from yellow to silver eel stage. Differentiation from intersexual gonads to testes took place after 2 years and was synchronized with metamorphosis. Differentiated ovaries, however, were already present in 2 + yellow eels indicating that expression of female gonadal sex was not synchronized with metamorphosis. Body length and eye size were used to assess the ontogenetic stage of individual fish. The majority of female yellow eel and male and female silver eel of the experimental fish could be identified using these external morphological characteristics. Intersexual yellow eel could not be identified with this method. Intersexual and female yellow eel reached the silver stage at ages of 2–3 and 3–4 years and at body weight ranges of 78–410 and 309–830 g, respectively, implying that metamorphosis in captivity took place at earlier ages and at heavier body weights than in nature. Metamorphosis with respect to enlargement of eyes up to the size of mature silver eels occurred in the hatchery population without hormonal intervention. The testes of silver eels contained spermatogonia in mitotic arrest and the ovaries had oocytes, which were blocked in late prophase of meiosis. Body length and eye size were not suitable characteristics to assess sexual maturity in hatchery raised silver eel. Males and females continued to eat and grow after they became silvery whereas gonadal development gradually stopped. The relationship between age, length/weight, feed intake and metamorphosis (including integumental colour change, enlargement of eyes, degeneration of the alimentary tract, gonadal growth/maturation) as well as the role of gonadotropins are discussed.

To test for a residual maternal influence in the performance of androgenetic offspring in common carp, Cyprinus carpio, an experiment was conducted where three females and three isogenic males were used to produce nine androgenetic families. Eggs were irradiated with UV (175 mJ cm−2) and diploidy was induced using a heat shock (40 °C, 2 min). Yields of androgenetic diploids depended mainly on the egg donor. After raising the larvae for a period of 10 weeks, significant differences were observed regarding the frequency of normal and deformed androgenetic animals, also depending on the egg donor. It is hypothesized that the UV-irradiation causes damage in maternal RNA's, resulting in disturbed embryonic development.

Haploid androgenetic offspring (max. 53.9%) were obtained after UV-irradiating eggs using dosages of 150–300 mJ/cm 2 (duration: 135 to 270 s at an intensity of 1.1 mW/cm 2 ). During irradiation, eggs were stirred in a synthetic ovarian fluid to assure a homogenous irradiation of all pronuclei, without activating the eggs. Androgenetic origin was checked using a recessive colour marker (“blond”). Absence of black fry (dominant trait) in the offspring confirmed inactivation of the female genome. Besides androgenetic haploids, few biparental diploids hatched at the optimal dose (250 mJ/cm 2 ). Diploidy could be restored by heat shocks (40.0°C, 2 min) applied at different times after fertilization (26, 28 and 30 min). Yields of putative androgenetic diploids ranged from 7.2 to 18.3% at the time of hatching. Also, survival of the larvae 24 days after hatching was high (78–89% of the initial number). After testing the homozygous nature of the fish, they will be used to generate isogenic strains.

A comparative study of larval growth in the different species of the genus Clarias in different regions revealed that in spite of strong differences in egg and larval size, the growth potential was quite similar. Differences in growth between species and regions were strongly correlated with the applied feeding strategy, the applied growth parameter and period of measuring. To evaluate feeding strategies, standardized procedures for measuring growth are needed. Recalculating the results of the mentioned study according to these standardized procedures revealed that both in India and Indonesia, the growth rate of C. batrachus larvae was too low. This is probably due to low feeding levels in India and premature weaning in Indonesia. The basic biology and nutritional physiology of the larvae of C. gariepinus have been studied extensively. At the start of exogenous feeding, the larvae have an advanced digestive system with a functional pancreas, liver and nutrient absorption capabilities, but lack a functional stomach. The advanced digestive system is further exemplified by the development of the enzymatic complex. Probably because of the rapid development of the digestive system, feeding live food organisms is mostly practiced for a few days only and is soon replaced by wet and/or dry diets. It is hypothesized that the requirement of live food or specific larval diets during the first days of exogenous feeding is related to the absence of pepsin digestion during this period. Further optimization of the feeding strategies should focus on this conclusion.