Influence of temperature on muscle fibre hyperplasia and hypertrophy in larvae of blackspot seabream,Pagellus bogaraveo

The effects of temperature on Pagellus bogaraveo muscle cellularity were investigated, by morphometry, throughout the endogenous feeding stage, testing two rearing temperatures: 14 and 18 °C. The following parameters were estimated in transversal body sections at post‐opercular and post‐anal body levels: the total cross‐sectional muscle area, the total number of fibres and the mean cross‐sectional fibre area. At hatching, no significant influence of the temperature was observed on the morphometric parameters measured in the white muscle. At mouth opening, an increase in the number of post‐opercular white fibres was promoted by the highest temperature. During embryonic development, the red muscle fibre number in the post‐anal part of the larvae increased with higher temperature, but it appears that the difference was no longer present at mouth opening. An increase in the fibre area and in the total cross‐sectional area of red muscle at the post‐anal level was promoted by a 4 °C increase in the temperature during the vitelline phase. In conclusion, the axial musculature of blackspot seabream embryos/larvae reacted differently to temperature influence according to the body location, strongly supporting the need to look at and account for different body locations when evaluating muscle cellularity in fish, namely in growth/aquaculture‐related studies.     

Growth and nutrient utilisation of blackspot seabream (<Emphasis Type="Italic">Pagellus bogaraveo</Emphasis>) under different feeding regimes

The growth and nutrient utilization of blackspot seabream was studied under self-feeding or hand feeding for 90 days. Groups of 31 fish with an initial body weight of 24 g were fed either by hand two times a day (09:00, and 18:00 h) to apparent satiety or by self-feeders. The 90 days of the feeding trial included two periods: an adaptation period (30 days) required to achieve a constant number of feed demands per day and a subsequent experimental period (60 days). Final body weight and daily growth index were unaffected by the feeding regimes. However, the marked reduction in voluntary feed intake associated with similar nutrient gain on the self-fed group resulted in improved nutrient efficiency and in subsequent increased protein, lipid and energy retentions compared to fish hand-fed at set hours. The self-fed group displayed depressed malic (<62%) and fatty acid synthetase (<35%) activities as well as reduced triacylglycerol plasma levels, which correlated positively with feed intake and, at some extent, with fish lipid content. These results indicate the ability of blackspot seabream to adjust their lipid metabolism according to fish feeding rhythm. No effect of feeding method was however observed on glycolytic hepatic activities or on glucose, cholesterol and insulin plasma levels. Self-feeders led to similar growth (DGI, 1.4–1.5) but better efficiency (FCR, 1.0 vs. 1.5), and hence, can be regarded as a helpful tool to optimize feed distribution according to this species natural rhythm. The maximal number of demands occurring between 20:00 and 21:00 h (dusk/sunset), together with the fact that 61% of the feed demands took place during the night, reveals a preferential crepuscular/nocturnal feeding pattern of this species.     

Muscle development and body growth in larvae and early post-larvae of shi drum, Umbrina cirrosa L., reared under different larval photoperiod: muscle structural and ultrastructural study

Shi drum specimens were maintained under four different photoperiod regimes: a natural photoperiod regime (16L:8D), constant light (24L), equal durations of light and dark (12L:12D) and a reduced number of daylight hours (6L:18D) from hatching until the end of larval metamorphosis. Specimens were then kept under natural photoperiod conditions until 111 days post-hatching. Muscle and body parameters were studied. During the vitelline phase, there was little muscle growth and no photoperiod effects were reported; however, a monolayer of red muscle and immature white muscle fibres were observed in the myotome. At hatching, external cells (presumptive myogenic cells) were already present on the surface of the red muscle. At the mouth opening, some presumptive myogenic cells appeared between the red and white muscles. At 20 days, new germinal areas were observed in the apical extremes of the myotome. At this stage, the 16L:8D group (followed by the 24L group) had the longest body length, the largest cross-sectional area of white muscle and the largest white muscle fibres. Conversely, white muscle hyperplasia was most pronounced in the 24L group. Metamorphosis was complete at 33 days in the 24L and 12L:12D groups. At this moment, both groups showed numerous myogenic precursors on the surface of the myotome as well as among the adult muscle fibres (mosaic hyperplastic growth). The 16L:8D group completed metamorphosis at 50 days, showing a similar degree of structural maturity in the myotome to that described in the 24L and 12L:12D groups at 33 days. When comparing muscle growth at the end of the larval period, hypertrophy was highest in the 16L:8D group, whereas hyperplasia was higher in the 24L and 16L:8D groups. At 111 days, all groups showed the adult muscle pattern typical of teleosts; however, the cross-sectional area of white muscle, white muscle fibre hyperplasia, body length and body weight were highest in the 24L group, followed by the 12L:12D group; white muscle hypertrophy was similar in all groups. Larval survival was higher under natural photoperiod conditions compared to all the other light regimes.     

Digestive enzyme activity at different developmental stages of blackspot seabream, Pagellus bogaraveo (Brunnich 1768)

Blackspot seabream, Pagellus bogaraveo (Brunnich), has been identified as a potential species to diversify European aquaculture production. Although rearing aspects have been widely investigated, little information exists on the nutritional requirements for this species. The aim of this study was to build up information on the activity of digestive enzymes at certain developmental stages of blackspot seabream in order to understand the nutritional needs of larvae and post larvae. Fish larvae were reared from hatching to 55 days after hatching (dah), and the feeding plan consisted in rotifers (5–35 dah), Artemia naupli (30–35 dah) metanaupli (35–45) and Gemma microdiet (45–55 dah). At 7, 11, 21, 45 and 55 days after hatching (dah), pooled samples of fish larvae were collected for analysis of trypsin, amylase, lipase, alkaline phosphatase and leucine–alanine peptidase activity. Up to 21 dah, the whole larvae body was used for enzymatic analysis, whereas in older larvae only the dissected abdominal cavity was used. Blackspot seabream body dry weight growth was exponential, increasing from 60 μg at 5 dah to 30±9.7 mg at 55 dah. Amylase specific activity decreased significantly during development, exhibiting at 11 dah (0.6 U mg^?1 protein) an average value 2.7 times lower than at 7 dah, and remaining stable between 45 and 55 dah (0.7 U mg protein^?1). Trypsin specific activity remained constant until 21 dah (between 38 and 44 mU mg protein^?1), which could be related to the larvae feeding regime. At later stages of development, lipase‐specific activity exhibited a significant increase (P<0.05), being three times higher at 55 dah (8 U mg protein^?1) than at 45 dah. The total activity of the studied digestive enzymes increased significantly during larval development (until 21 dah), whereas afterwards only lipase and leucine–alanine peptidase increased significantly between 45 and 55 dah. The pattern of digestive enzymes activity was related to organogenesis and the type of food used at different developmental stages.     

Weaning of the wedge sole <Emphasis Type="Italic">Dicologoglossa cuneata</Emphasis> (Moreau): influence of initial size on survival and growth

The weaning phase can be decisive in fish-culture viability. In this work, the relationship between the initial size and weaning success has been studied in wedge sole (Dicologoglossa cuneata). For each age (30, 50, and 70 days after hatching, DAH), two to three sizes were selected, and all were put on the same feeding schedule for 20 days. Each batch (three replicates) was sampled at 1, 10, and 20 days. Specific growth rate (SGR) and survival were compared at the end of the co-feeding period, after 10 days on dry feed only. The best results for survival and growth were found with the smallest larvae, and vice versa. The SGRs and survival rates recorded during the co-feeding period were higher (0.8–15.6 day⁻¹ and 68.3–97.8%) than those from the dry-food phase (0.9–4.7 day⁻¹ and 56.3–66.7%). Successful weaning (survival = 65% and SGR = 9.3 day⁻¹) is possible with 30 DAH larvae (7.6–8.1 mm and 3.9–4.6 mg). In conclusion, the most effective weaning would be possible at 30 DAH, implying significant Artemia savings (25–50%).     

Histological development of digestive tract in discus, <Emphasis Type="Italic">Symphysodon</Emphasis> spp. larvae

The present study provides information on the histomorphological development of digestive system of discus, Symphyosodon spp., larvae during the first month of life. Discus larvae are altricial at hatching, with an undifferentiated digestive tract and a large yolksac, which is completely consumed within 7 days. The mouth opens 3 days after hatching (DAH) and the larvae starts feeding on AF Artemia at 4 DAH when offered. At 3 DAH the digestive tract is differentiated with distinct esophagus, stomach anlage, and mid- and hindguts. At 5 DAH, discus larvae is an active feeder, equipped with partly developed jaws and ossified gill arches and an inflated swim bladder. The liver and pancreas are present and supranuclear inclusion vacuoles (SIV) appear in the hindgut for the first time. Gastric glands in stomach were first observed 7 DAH and proliferated by 11–13 DAH. SIV were a common feature in the midgut and hindgut epithelium until 15–23 DAH. Therefore, exclusive use of artificial diets should be postponed until 2–3 weeks after hatching.     

Nutritional stimuli of gilthead seabream (Sparus aurata) larvae by dietary fatty acids: effects on larval performance,gene expression and neurogenesis

The concept of nutritional programming raises the interesting possibility of directing specific metabolic pathways or functions in juvenile fish, for example, to improve the use of substitutes to fishmeal and oil, and hence to promote sustainability in aquaculture. The aim of the study was to determine effects of early nutritional stimuli of gilthead seabream larvae and check if nutritional programming of gilthead sea bream is possible between 16 days post hatching (dph) and 26 dph. A trial was conducted to determine the effects of early nutritional stimuli of gilthead seabream larvae. Five experimental microdiets (pellet size <250 μm) were formulated containing five different proportions of a marine lipid source rich in long‐chain polyunsaturated fatty acids (LC‐PUFA) and two vegetable lipid sources rich in linolenic and linoleic acids. The results of this study demonstrate that dietary n‐3 LC‐PUFA levels increased larval growth and survival affecting Δ6 desaturase gen (fads2) expression and retinal neurons density. However, the high mortalities obtained along on‐growing in fish fed low n‐3 LC‐PUFA at 16 dph constrained the feasibility of nutritional programming of gilthead seabream during this late developmental window and needs to be further investigated.     

Organogenesis of exocrine pancreas in sharpsnout sea bream (<Emphasis Type="Italic">Diplodus puntazzo</Emphasis>) larvae: characterization of trypsin expression

The ontogeny and differentiation stages of digestive systems related with trypsin expression in larvae of sharpsnout sea bream, Diplodus puntazzo, were investigated from hatching to 40 DAH (days after hatching), and total lengths and weights of larvae were determined. Histologic and enzymatic techniques were used to explain the functional development of the pancreas including trypsin activity. The pancreas was identified as a compact structure located in the region slightly posterior to the liver. At 3 DAH, first anus and then mouth opened. Incipient pancreas secretion polyhedral cells could be first observed as zymogen granules. During larval metamorphosis, the pancreas became diffuse, spreading throughout the mesentery in proximity to the stomach, the anterior intestine and the pyloric caeca. The specific activity of trypsin (42.54 ± 6.8 mU/mg protein⁻¹) was found as early as after hatching at larvae size of 2.87 ± 0.34 mm at 0 DAH. Activity further increased until 10 DAH, especially after exogenous feeding. The highest trypsin activity was detected at 25 DAH as 119.26 ± 11.6 mU/mg protein⁻¹. It is concluded that exocrine pancreas organogenesis is the main critical step in the development of digestive system that results in zymogen granules accumulation and increased trypsin activity.     

Effects of the timing of initial feeding on growth and survival of loach (<Emphasis Type="Italic">Misgurnus anguillicaudatus</Emphasis>) larvae

The effects of delayed first feeding on growth and survival and starvation on the point-of-no-return of loach Misgurnus anguillicaudatus larvae were studied by evaluating morphometric characteristics under controlled conditions. Larvae began to feed exogenously at 3 days after hatching (DAH) and the PNR occurred between 9 and 10 DAH at 23 ± 1.0°C. The experimental design included a conventional feeding regime with initial feeding from 3 DAH as a control, delayed first feeding for 4, 5 and 6 DAH. Morphometric characteristics (head depth, body depth, eye diameter, mouth diameter, musculature height, total length and yolk sac volume) were evaluated under different initial feeding time (3, 4, 5 and 6 days after hatching). Loach larvae initiated first feeding at 4, 5 and 6 days after hatching achieved comparatively lesser growth performance in all morphometric characteristics than that of 3 days at the end of the experiment. By day 6, significant differences were observed between 3 and 6 days initial feeding larvae for all morphometric characteristics except eye diameter and mouth diameter. Similarly, significant differences were noticed between 3 and 5 days initial feeding. However, there were no significant differences in head depth, body depth, eye diameter, mouth diameter, and total length between 3 and 4 days initial feeding until 12 DAH. After 15 days rearing, significant differences in all morphometric characteristics appeared between 3 and 4 days initial feeding and followed to the end of the experiment. It was also observed that the yolk absorption in loach larvae was completed by 6 days irrespective of the differences in the initial feeding. The yolk volume of 4 and 5 DAH larvae initiated first feeding at 3 days (0.0125 ± 0.0015; 0.0077 ± 0.0009 mm³) had significant differences compared with yolk volume of larvae initiated first feeding at 4 days (0.0081 ± 0.0011; 0.0039 ± 0.0004 mm³), 5 days (0.0079 ± 0.0010; 0.0017 ± 0.0002 mm³) and 6 days (0.0082 ± 0.0011; 0.0016 ± 0.0001 mm³). Survival rates of four treatments were estimated daily for 30 days and significant differences were observed between the treatments at the end of the experiment. The final survival rate was higher when the loach larvae initiated feeding at 3 days (75.9%) when compared with 4 days (31.8%), 5 days (14.5%) and 6 days (6.4%). The present study suggests that the first feeding of loach larvae should be initiated at 3 DAH for achieving better growth and survival or else bad growth performance will engender if the first feeding is delayed.     

Effects of Feeding Levels on Growth Response,Body Composition,and Energy Expenditure in Blackspot Seabream,Pagellus bogaraveo,Juveniles

Although reported as a slow growing species, to date information regarding nutritional dynamics of blackspot seabream, Pagellus bogaraveo, is limited, especially during ongrowing phase. This study determined the effects of different feeding levels on growth response, body composition, and energy expenditure of blackspot seabream juveniles (initial weight = 64 g). Fish were allocated in nine indoor tanks (15 fish per tank) and fed a basal diet twice daily at increasing levels (2.5, 3.3, and 4.1 g feed/kg^0.8/d) for 138 d. Growth response decreased with decreasing feeding levels, but only at 2.5 g feed/kg^0.8/d (P < 0.001). Differences in growth rates were positively correlated with body lipid content (r² = 0.90). Net protein utilization decreased with each increase in feeding level. The energy requirement for zero growth (maintenance) and the estimated energy loss on starvation were calculated as 22.6 and 17.9 kJ/kg^0.8/d, respectively. The energy cost for protein and lipid gain was calculated as 3.8 and 1.4 kJ/kJ energy deposited, respectively. This study suggested that blackspot seabream may grow slower than the other close‐related species because of its higher demand of dietary protein energy in favor of lipid deposition.     

Applying the ontogeny of digestive enzyme activity to guide early weaning of pigfish,Orthopristis chrysoptera (L.)

Weaning marine fish larvae from live prey to a dry microdiet is an important step towards optimizing the commercial production, but early weaning is constrained by the lack of sufficient digestive enzymes at first feeding. This study quantified the activity of five digestive enzymes throughout the larval period of pigfish (Orthopristis chrysoptera [L.]) to assess ontogenetic changes in digestive abilities, and then trials were conducted that determined the optimal time for weaning. The activity of all digestive enzymes was low or undetectable at first feeding (3 days post hatching, dph; 2.5 mm standard length, SL). A substantial increase in activity occurred at 5.7 mm SL (17 dph), 6.9 mm SL (21 dph), 7.7 mm SL (23 dph), 8.4 mm SL (25 dph) and 11.2 mm SL (30 dph) for bile salt‐dependent lipase, trypsin, chymotrypsin, amylase and acid protease respectively. During the weaning experiment, larvae were co‐fed live prey and microdiet beginning 15 dph (4.8 mm SL). Live prey was withdrawn from the diet at 24, 28, 32 or 36 dph, with the control receiving live prey and microdiet throughout (to 43 dph). There were no significant differences in mean final SL among treatments, but survival was significantly lower when larvae were weaned at 24 dph compared to 32–43 dph. Based on the digestive enzyme activity and survival, weaning larval pigfish at 32 dph (11.7 mm SL) when reared at 24°C is recommended.     

Ontogeny of the digestive system in hatchery produced Beluga (Huso huso Linnaeus, 1758); a comparative study between Beluga and genus Acipenser

The study on histological characteristics of the digestive system of Beluga (Huso huso) was conducted from hatching until 50 days posthatching at 16.5 °C. Development of the digestive system in this species followed the general pattern described for other Acipenserids, although there were differences in the timing of organ development among species. At hatching, the mouth was opened and digestive system was represented by a gastric cavity filled with yolk and lined by endodermal cells, and a partially differentiated hindgut. Gastric glands started to differentiate at 46.5 degree‐ days posthatching (ddph), the earliest appearance time among sturgeon fishes studied to date. At the onset of exogenous feeding (144.9 ddph), yolk sac reserves were not completely depleted in the stomach, suggesting a period of mixed nutrition. The complete development of the digestive system was not accomplished until 235.2 ddph when it showed all histomorphological features typical of juvenile specimens. According to histological results, it seems advisable to start co‐feeding H.huso larvae with inert diets at the onset of exogenous feeding, because exocrine pancreas and glandular stomach are fully differentiated, although the complete substitution of live prey by inert feed is not recommended until 235 ddph.     

Plant protein and vegetable oil‐based diets modulate gilthead seabream (Sparus aurata) muscle biochemical status and proteolytic enzymes at an early postmortem stage

A 70 days feeding trial was carried out on juvenile gilthead seabream (Sparus aurata) (mean initial body weight: 362 ± 59 g) to evaluate whether the simultaneous replacement of fishmeal and fish oil by vegetable ingredients affected the muscle's proteolytic potential. Fish were fed either a fishmeal/fish oil diet, a fishmeal/vegetable oil diet, a plant protein/fish oil or a plant protein/vegetable oil diet. The use of dietary plant proteins resulted in lower sulphated glycosaminoglycans' content and lower glycogen phosphorylase activity in the muscle, while both plant proteins and vegetable oil increased muscle pH and reduced cathepsin B activity. Our results indicate that high replacement of fish meal and/or fish oil with plant ingredients has a significant impact on early postmortem metabolic processes and proteolytic potential of seabream muscle, especially when replacing both fish meal and fish oil.     

Effect of storage time and cryoprotectant concentrations on the fertilization rate and hatching rate of cryopreserved sperm in red seabream (Pagrus major Temminck & Schlegel, 1843)

This study examined the effects of storage time and cryoprotectant concentrations on the post‐thaw sperm of red seabream, Pagrus major. Sperm treated with 12%, 15%, 18% and 21% DMSO were cryopreserved for 10, 30, 60 and 360 days, and fertilization and hatching rates were analysed. For all groups, there were no differences in the fertilization rates and hatching rates between sperm cryopreserved for <60 days and fresh sperm (98.8±0.8%, 96.4±1.3%). However, for sperm cryopreserved for 360 days, both fertilization rates (88.6±3.0% to 7.0±1.9%) and hatching rates (79.4±7.2% to 3.3±0.8%) decreased drastically. Furthermore, the cryoprotectant concentrations affected sperm quality significantly (P<0.05). When cryopreserved for 360 days, sperm treated with 15% DMSO obtained the best results compared with other concentrations. We suggest that 15% DMSO may be an effective cryoprotectant for long‐term sperm cryopreservation of red seabream.     

Ontogenetic development of digestive enzymes and effect of starvation in miiuy croaker <Emphasis Type="Italic">Miichthys miiuy</Emphasis> larvae

The ontogenetic development of the digestive enzymes amylase, lipase, trypsin, and alkaline phosphatase and the effect of starvation in miiuy croaker Miichthys miiuy larvae were studied. The activities of these enzymes were detected prior to exogenous feeding, but their developmental patterns differed remarkably. Trypsin activity continuously increased from 2 days after hatching (dah), peaked on 20 dah, and decreased to 25 dah at weaning. Alkaline phosphatase activity oscillated at low levels within a small range after the first feeding on 3 dah. In contrast, amylase and lipase activities followed the general developmental pattern that has been characterized in fish larvae, with a succession of increases or decreases. Amylase, lipase, and trypsin activities generally started to increase or decrease at transitions from endogenous to exogenous feeding or diet changes, suggesting that these enzymatic activities can be modulated by feeding modes. The activities of all the enzymes remained stable from 25 dah onwards, coinciding with the formation of gastric glands and pyloric caecum. These results imply that specific activities of these enzymes underwent changes due to morphological and physiological modifications or diet shift during larval development but that they became stable after the development of the digestive organs and associated glands was fully completed and the organs/glands functioned. Trypsin and alkaline phosphatase were more sensitive to starvation than amylase and lipase because delayed feeding up to 2 days after mouth opening was able to adversely affect their activities. Enzyme activities did not significantly differ among feeding groups during endogenous feeding; however, all activities were remarkably reduced when delayed feeding was within 3 days after mouth opening. Initiation of larvae feeding should occur within 2 days after mouth opening so that good growth and survival can be obtained in the culture.     

Dietary protein, growth, nutrient utilization and body composition of juvenile blackspot seabream, Pagellus bogaraveo (Brunnich)

When considering new marine species for Mediterranean aquaculture, blackspot seabream emerges as a potential candidate. However, there are scarce data on the nutritional requirements and optimal growth conditions of this species. A 12‐week feeding trial was conducted to evaluate the effects of dietary protein on growth, nutrient utilization and body composition of blackspot seabream (23 g). Five isolipidic diets (12%) with graded levels of protein (20–60%) were distributed, twice a day, to duplicate groups of fish, until satiation. Growth increased significantly with increasing dietary protein up to 40%, but higher protein levels induced a similar daily growth index (1.4). Feed conversion ratio (FCR) decreased with increasing levels of dietary protein (4.2–1.6). No significant differences were detected in protein of whole body blackspot seabream among treatments, but fat percentage decreased with increasing dietary protein. Dry matter and energy digestibility showed a concomitant increase with the reduction in dietary wheat meal, attaining maximal values with high protein diets. These results suggested that the most favourable values for growth and FCR are obtained with diets containing 40% protein. However, the excessive lipid deposition reveals that more nutritional studies are necessary before the species can be established in aquaculture.     

Development of digestive enzyme activity in larvae of spotted sand bass <Emphasis Type="Italic">Paralabrax maculatofasciatus</Emphasis>. 1. Biochemical analysis

Spotted sand bass Paralabrax maculatofasciatus is a potential aquaculture species in Northwest Mexico. In the last few years it has been possible to close its life cycle and to develop larviculture technology at on pilot scale using live food, however survival values are low (11%) and improvements in growth and survival requires the study of the morpho-physiological development during the initial ontogeny. In this research digestive activity of several enzymes were evaluated in larvae, from hatching to 30 days after hatching (dah), and in live prey (rotifers and Artemia), by use of biochemical and electrophoretic techniques. This paper, is the first of two parts, and covers only the biochemical analysis. All digestive enzyme activities were detected from mouth opening; however the, maximum activities varied among different digestive enzymes. For alkaline protease and trypsin the maximum activities were detected from 12 to 18 dah. Acid protease activity was observed from day 12 onwards. The other digestive enzymes appear between days 4 and 18 after hatching, with marked fluctuations. These activities indicate the beginning of the juvenile stage and the maturation of the digestive system, in agreement with changes that occur during morpho-physiological development and food changes from rotifers to Artemia. All enzymatic activities were detected in rotifers and Artemia, and their contribution to enhancement the digestion capacity of the larvae appears to be low, but cannot be minimised. We concluded that the enzymatic equipment of P. maculatofasciatus larvae is similar to that of other marine fish species, that it becomes complete between days 12 and 18 after hatching, and that it is totally efficient up to 25 dah.     

Fish oil replacement by different microalgal products in microdiets for early weaning of gilthead sea bream (Sparus aurata,L.)

The aim of this study was to determine if algal products rich in DHA or ARA are able to completely replace fish oil in microdiets for marine fish larvae, gilthead seabream and if extra supplementation with EPA may further enhance larval performance. For that purpose, 20 day‐old gilthead seabream larvae of 5.97 ± 0.4 mm mean total length and 0.12 ± 0.001 mg mean dry body weight were fed with five microdiets tested by triplicate: a control diet based on sardine oil; a diet containing AquaGrow^® DHA (diet DHA) to completely substitute the sardine oil; a diet containing AquaGrow^® ARA (diet ARA); a diet containing both products, AquaGrow^® DHA and AquaGrow^® ARA to completely substitute the fish oil; and, a diet containing both products, AquaGrow^® DHA and AquaGrow^® ARA, together with an EPA source. Temperature, air and salinity activity tests were also performed to detect larval resistance to stress. At the end of the experiment, final survivals did not differ among groups. The microorganism produced DHA was able to completely replace fish oil in weaning diets for gilthead seabream without affecting survival, growth or stress resistance, whereas the inclusion of microorganism produced ARA did not improve larval performance. Moreover, addition of EPA to diets with total replacement of fish oil by microorganism produced DHA and ARA, significantly improved growth in terms of body weight and total length. The results of this study denoted the good nutritional value of microorganisms produced DHA as a replacement of fish oil in weaning diets for gilthead seabream, without a complementary addition of ARA. However, dietary supplementation of EPA seems to be necessary to further promote larval performance.     

Is it possible to differentiate between environmental and fishery effects on abundance‐biomass variation? A case study of blackspot seabream (Pagellus bogaraveo) in the Strait of Gibraltar

We assessed the potential for simulation and modelling of the blackspot seabream (Pagellus bogaraveo) population in the Strait of Gibraltar to discriminate the environmental effects of fishery impacts. A discrete biomass–abundance dynamic model was implemented to obtain a simulated monthly time series of blackspot seabream biomass. On this simulated time series, autoregressive integrated moving average (ARIMA) models were fitted. The best ARIMA fit provided a significant correlation of 0.76 and persistence index higher than 0.85. The proportion of variance non‐explained by the ARIMA models was correlated with a time series of sea surface temperature (SST) and North Atlantic Oscillation (NAO). The analysis of global, annual and winter correlation between the proportion of variance not explained by the ARIMA models and environmental variables showed that significant associations were not detected over the full time series. Our analysis therefore suggests that overexploitation is the main factor responsible for the commercial depletion of blackspot seabream in the Strait of Gibraltar.     

Partial sequence characterization and ontogenetic expression of genes involved in lipid metabolism in the tropical gar (Atractosteus tropicus)

The tropical gar (Atractosteus tropicus) is an economically and socially important freshwater fish species from southeastern Mexico, with high aquaculture potential. This study determined the expression of acetyl‐CoA carboxylase (ACC1), fatty acid synthase (FAS), and carnitine palmitoyl transferase 1 (CPT1C) in several tissues of adult specimens, as well as in early ontogeny stages of tropical gar. Partial sequences of all genes were obtained for brain, liver, muscle, mesenteric adipose, kidney, testis, gill, and heart in males. Specific qPCR primers were used to compare the expression between tissues and also during larviculture, starting with embryos (0 days after hatching, [DAH]), and at 5, 10, 15, 20, 25, and 30 DAH. Mesenteric adipose tissue, liver, and muscle showed maximum FAS, ACC1, and CPT1C expression. Additionally, the highest expression of lipogenic genes was detected in embryos (0 DAH), decreasing between 5 and 10 DAH, then increasing at 15 DAH, and ultimately decreasing again from 20 DAH onwards. Based on these results, it can be concluded that genes encoding ACC1, FAS, and CPT1C are expressed differentially across tissues of tropical gar adults, and are regulated during larval development.