Digestive enzyme profiles reveal digestive capacity and potential energy sources in fed and starved spiny lobster (Jasus edwardsii) phyllosoma larvae

The impact of starvation on the digestive enzyme (protease, trypsin, lipase and amylase) activities of Stage I and IV Jasus edwardsii phyllosoma larvae was used to identify the nutrients metabolised or conserved during food deprivation, highlighting the most critical energy reserves. Protease activities increased significantly in both Stages I and IV phyllosoma, suggesting that protein catabolism provided energy during food deprivation. Lipase activity decreased significantly in starved Stages I and IV larvae indicating that lipid may be spared for fuelling later developmental moults. The use of protein, while sparing lipid, may provide immediate energy but not at the expense of long-term lipid energy stores which are known to be important during their lengthy larval phase. The preferential use of protein during short-term periods of starvation suggests that particular attention must be given to providing sufficient protein in artificial diets at all times. Amylase activity in starved Stage I larvae was lower than in fed animals, suggesting that the starved animals are not gaining sufficient carbohydrate. However, amylase activity was similar in fed and starved Stage IV larvae, possibly due to the catabolism of accumulated glycogen stores that were not sufficiently developed in Stage I animals.     

Metabolic fate of yolk fatty acids in the developing king penguin embryo

This study examines the metabolic fate of total and individual yolk fatty acids (FA) during the embryonic development of the king penguin, a seabird characterized by prolonged incubation (53 days) and hatching (3 days) periods, and a high n-3/n-6 polyunsaturated FA ratio in the egg. Of the approximately 15 g of total FA initially present in the egg lipid, 87% was transferred to the embryo by the time of hatching, the remaining 13% being present in the internalized yolk sac of the chick. During the whole incubation, 83% of the transferred FA was oxidized for energy, with only 17% incorporated into embryo lipids. Prehatching (days 0-49), the fat stores (triacylglycerol) accounted for 58% of the total FA incorporated into embryo lipid. During hatching (days 49-53), 40% of the FA of the fat stores was mobilized, the mobilization of individual FA being nonselective. At hatch, 53% of the arachidonic acid (20:4n-6) of the initial yolk had been incorporated into embryo lipid compared with only 15% of the total FA and 17-24% of the various n-3 polyunsaturated FA. Similarly, only 32% of the yolk's initial content of 20:4n-6 was oxidized for energy during development compared with 72% of the total FA and 58-66% of the n-3 polyunsaturated FA. The high partitioning of yolk FA toward oxidization and the intense mobilization of fat store FA during hatching most likely reflect the high energy cost of the long incubation and hatching periods of the king penguin. The preferential partitioning of 20:4n-6 into the structural lipid of the embryo in the face of its low content in the yolk may reflect the important roles of this FA in tissue function.     

Some effects of early starvation on the survival and development of barnacle nauplii,Balanus improvisus (Darwin)

Newly hatched stage I–II nauplii of Balanus improvisus (Darwin) were “totally starved” (until death) or “partially starved” for the first 48 h and 96 h of their development. Daily mortality and molting were monitored throughout larval development in both starved and fed control groups. Fed control animals exhibited a largely synchronous molting pattern with instars of equal duration. Total starvation suppressed molting beyond stage II; 50% mortality occurred in ≈4 days at both 15 and 21 °C, while longest survival time was 7 days at 15 °C and 6 days at 21 °C. At 15 °C, partially starved nauplii retained the ability to complete naupliar development but at a slower overall rate and with increased mortality relative to controls. These effects were more pronounced in the 96-h group. Increased mortality of stage VI nauplii was evident in both partially starved groups (7.1% for 48 h, 18.8% for 96 h) relative to unstarved controls (3.1%). Stage II nauplii exhibited little resistance to starvation and survival potential may have decreased as soon as 24 h.     

Evaluation of the fatty acid profiles of two fairy shrimp species,Branchipus pasai Cottarelli, 1969 and Chirocephalus kerkyrensis Pesta, 1936 (Crustacea,Anostraca) fed different diets

Laboratory cultured fairy shrimps Branchipus pasai and Chirocephalus kerkyrensis, fed on an alga (Selenastrum capricornutum), a yeast (Saccharomyces cerevisiae) and an HUFA enriched yeast (Lansy PZ, produced by Artemia Systems, Gent, Belgium) were evaluated for their fatty acid (FA) profiles and total lipid content in order to obtain information on species differences in food conversion. The results indicate significant qualitative and quantitative differences (P < 0.001) in FA profiles both of feed and of fairy shrimp species. Among the three different diets, an appreciable lipid amount was recorded in the alga, as compared with baker‘s yeast which showed the poorest lipid content. The algal fatty acid profile showed adequate amounts of the EFA 18:2n6, 18:3n3 and20:5n3 (the most meaningful for aquacultural purposes). The enriched yeast was characterised by a considerable total lipid amount and by the presence of all the EFA. The overall amounts of fatty acids in the fairy shrimps correlated well with their levels in the feed(r > 0.9, P <0.001). An exception was noted in then-3 HUFA (highly unsaturated fatty acids) and n-6acids, among the individuals fed on yeast. Highly significant differences (P < 0.001) were also noted between the two fairy shrimp species fed on the same food type. When fed enriched yeast, both B. pasai and C. kerkyrensis profiles roughly reflected diet composition. However, when fed on algae or baker‘s yeast, the two species, though to different extents, exhibited higher EFA levels than those recorded in the food. This seems to support the existence of a partial FA bioconversion capacity in fairy shrimps previously noted in the case of the brine shrimp Artemia. This revised version was published online in August 2006 with corrections to the Cover Date.     

Effects of two Avtemia diets with different contents of polyunsaturated fatty acids on the lipid composition of larvae of Atlantic herring (Clupea harengus

Atlantic herring larvae ( Clupea harengus ) were fed two enriched Artemia diets with different contents of (n-3) highly unsaturated fatty acids (HUFA), one containing low levels of 20: 5(n-3) and no 22: 6(n-3), the other containing substantial levels of both 20: 5(n-3) and 22: 6(n-3). After 30 days of culture, fatty acid compositions of lipid classes in the heads, bodies and eyes of the larvae were analysed. Fish fed Artemia with the low (n-3) HUFA diet lacking 22: 6(n-3) had lower amounts of total (n-3)HUFA and, in particular, of 22:6(n-3) in individual phospholipids and total neutral lipids of heads, bodies and eyes as compared to fish fed Artemia with high levels of (n-3)HUFA. The amount of 22: 6(n-3) in the fatty acids of phosphatidyl-ethanolamine of eyes was particularly susceptible to dietary depletion. The implications of these findings are discussed, particularly in relation to dietary requirements for 22: 6(n-3) during development of neural tissue in predatory fish iarvae.     

不同饵料对卤虫生长、总脂含量及脂肪酸组成的影响

为了提高养殖卤虫的饵料营养价值,了解其不同生长阶段营养成分变化情况,采用单因子试验研究了8种饵料（三角褐指藻、小球藻、微绿球藻、酵母液、三角褐指藻＋小球藻＋微绿球藻、三角褐指藻＋酵母液、小球藻＋酵母液和微绿球藻＋酵母液）对卤虫生长、总脂含量及脂肪酸组成的影响,结果表明：不同饵料种类对卤虫生长、总脂含量及脂肪酸组成的影响显著（P＜0.05）,增长率,以三角褐指藻＋酵母液最优;总脂含量、以三角褐指藻最优（19.67%）,除酵母液外,与其它饵料相差不显著（P＞0.05）;脂肪酸组成效果,以微绿球藻组最优（EPA：18.01%,DNA：0.55%,（n-3）HUFA：19.08%）,与三角褐指藻组相差不大（P＞0.05）,显著高于其它各组（P＞0.05）.同时以三角褐指藻为饵料,研究了卤虫不同生长阶段（体长2、4、6、8、10 mm）总脂含量、脂肪酸组成变化,结果表明：卤虫体长2～10 mm总脂含量为14.27%～20.93%,随体长的增长降低;EPA、DHA及（n-3）HUFA的含量,均随体长的增长降低,EPA含量为：10.47%～20.77%,DNA含量为：0～0.70%,（n-3）HUFA含量为：10.85%～22.01%.结论认为,卤虫以三角褐指藻或三角褐指藻＋酵母液为饵料培养营养价值最佳,其体长小于6 mm营养价值较佳.     

Dietary lipid quality and mitochondrial membrane composition in trout: responses of membrane enzymes and oxidative capacities

To examine whether membrane fatty acid (FA) composition has a greater impact upon specific components of oxidative phosphorylation or on overall properties of muscle mitochondria, rainbow trout (Oncorhynchus mykiss) were fed two diets differing only in FA composition. Diet 1 was enriched in 18:1n-9 and 18:2n-6 while Diet 2 was enriched in 22:6n-3. The FA composition of mitochondrial phospholipids was strongly affected by diet. 22:6n-3 levels were twice as high (49 %) in mitochondrial phospholipids of fish fed Diet 2 than in those fed Diet 1. 18:2n-6 content of the phospholipids also followed the diets, whereas 18:1n-9 changed little. All n-6 FA, most notably 22:5n-6, were significantly higher in fish fed Diet 1. Nonetheless, total saturated FA, total monounsaturated FA and total polyunsaturated FA in mitochondrial phospholipids varied little. Despite a marked impact of diet on specific FA levels in mitochondrial phospholipids, only non-phosphorylating (state 4) rates were higher in fish fed Diet 2. Phosphorylating rates (state 3), oxygen consumption due to flux through the electron transport chain complexes as well as the corresponding spectrophotometric activities did not differ with diet. Body mass affected state 4 rates and cytochrome c oxidase and F ₀ F ₁ ATPase activities while complex I showed a diet-specific effect of body mass. Only the minor FA that were affected by body mass were correlated with functional properties. The regulated incorporation of dietary FA into phospholipids seems to allow fish to maintain critical membrane functions even when the lipid quality of their diets varies considerably, as is likely in their natural environment.     

Dietary polyunsaturated fatty acids alter myocardial protein kinase C expression and affect cardioprotection induced by chronic hypoxia

We examined the influence of dietary fatty acid (FA) classes on the expression of protein kinase C (PKC) delta and epsilon in relation to the cardioprotective effects of chronic intermittent hypoxia (CIH). Adult male Wistar rats were fed a nonfat diet enriched with 10% lard (saturated FA [SFA]), fish oil (n-3 polyunsaturated FA [n-3 PUFA]), or corn oil (n-6 PUFA) for 10 weeks. After 4 weeks on the diet, each group was divided into two subgroups that were either exposed to CIH in a barochamber (7000 m, 8 hrs/ day) or kept at normoxia for an additional 5-6 weeks. A FA phospholipid profile and Western blot analysis of PKC were performed in left ventricles. Infarct size was assessed in anesthetized animals subjected to 20-min coronary artery occlusion and 3-hr reperfusion. CIH decreased the n-6/n-3 PUFA ratio in all groups by 23% independently of the initial value set by various diets. The combination of n-3 diet and CIH had a stronger antiarrhythmic effect during reperfusion than the n-3 diet alone; this effect was less pronounced in rats fed the n-6 diet. The normoxic n-6 group exhibited smaller infarctions (by 22%) than the n-3 group. CIH decreased the infarct size in n-3 and SFA groups (by 20% and 23%, respectively) but not in the n-6 group. Unlike PKC epsilon, the abundance of PKC delta in the myocardial particulate fraction was increased by CIH except for the n-6 group. Myocardial infarct size was negatively correlated (r=- 0.79) with the abundance of PKC delta in the particulate fraction. We conclude that lipid diets modify the infarct size-limiting effect of CIH by a mechanism that involves the PKC delta-dependent pathway.     

Maternal dietary n-3 fatty acids alter immune cell fatty acid composition and leukotriene production in growing chicks

The effect of feeding different amounts of n-6 and n-3 fatty acids (FA) to hens on immune tissue FA composition and leukotriene production of hatched chicks was investigated. Hens were fed diets supplemented with either 3.0% sunflower oil (Diet I), 1.5% sunflower+1.5% fish oil (Diet II), or 3.0% fish oil (Diet III) for 46 days. The hatched chicks were fed a diet containing C18:3n-3, but devoid of longer chain n-6 and n-3 FA, for 21 days. Spleen docosahexaenoic acid (DHA) content was higher in chicks from hens fed Diet III (P<0.05). The bursa content of arachidonic acid was lower in chicks hatched from hens fed Diet III (P<0.05), and the ratio of n-6 to n-3 FA was significantly higher in bursa of chicks hatched to hens fed Diet I (P<0.05). Eicosapentaenoic acid (EPA) and DHA contents were higher in bursa of chicks hatched from hens fed Diet III (P<0.05). Thrombocytes from chicks hatched to hens fed Diet III produced the most leukotriene B(5) (LTB(5)). The ratio of LTB(5) to LTB(4) concentrations was also highest (P<0.05) in chicks hatched to hens fed Diet III. These results indicate that modulating maternal dietary n-6 and n-3 FA may alter leukotriene production in chicks, which could lead to less inflammatory-related disorders in poultry.     

The ontogeny of physiological response to light intensity in early stage spiny lobster (Jasus edwardsii) larvae

Early stage phyllosoma larvae of the spiny rock lobster Jasus edwardsii were examined for swimming speed, feeding, oxygen consumption and nitrogen excretion as instantaneous performance indicators when exposed to different irradiance levels. Swimming speed was measured in recently hatched Stage I larvae while all other parameters were measured in larvae from hatch to mid-Stage V. The swimming speed of recently hatched Stage I phyllosoma increased logarithmically between light intensities of 2.9 x 10(14) and 1.8 x 10(16) quanta s(-1) cm(-2) indicating that, within this range, swimming activity was only suppressed at the lowest irradiance level. Larvae examined under dark (no light) conditions showed lower feed intake, oxygen consumption and nitrogen excretion than larvae under low (7.7 x 10(12) q s(-1) cm(-2)) and high (3.9 x 10(14) q s(-1) cm(-2)) light intensities, and this was a consistent pattern observed throughout development from hatch to Stage V. There was no difference in feeding, oxygen consumption and nitrogen excretion between larvae exposed to low and high light intensities. However, from mid-Stage I to mid-Stage V, the metabolic feeding efficiency (feed intake:oxygen consumption ratio) was consistently higher in larvae exposed to low light intensity than in phyllosoma assessed in the dark and under high irradiance. A light intensity of about 7.7 x 10(12) quanta s(-1) cm(-2) and no higher than 3.9 x 10(14) quanta s(-1) cm(-2) is recommended to stimulate feeding and optimise metabolic feeding efficiency in early larval stages of J. edwardsii.     

Interactive effects of maternal and weaning high linoleic acid intake on hepatic lipid metabolism,oxylipins profile and hepatic steatosis in offspring

Non-alcoholic fatty liver disease (NAFLD) has been described as a hepatic manifestation of the metabolic syndrome. When several studies correlated maternal linoleic acid (LA) intake with the development of obesity, only few links have been made between n-6 fatty acid (FA) and NAFLD. Herein, we investigated the influence of both maternal and weaning high LA intake on lipid metabolism and susceptibility to develop later metabolic diseases in offspring. Pregnant rats were fed a control-diet (2% LA) or a LA-rich diet (12% LA) during gestation and lactation. At weaning, offspring was assigned to one of the two diets, i.e., either maintained on the same maternal diet or fed the other diet for 6 months. Physiological, biochemical parameters and hepatic FA metabolism were analyzed. We demonstrated that the interaction between the maternal and weaning LA intake altered metabolism in offspring and could lead to hepatic steatosis. This phenotype was associated with altered hepatic FA content and lipid metabolism. Interaction between maternal and weaning LA intake led to a specific pattern of n-6 and n-3 oxylipins that could participate to the development of hepatic steatosis in offspring. Our findings highlight the significant interaction between maternal and weaning high LA intake to predispose offspring to later metabolic disease and support the predictive adaptive response hypothesis.     

Impact of feeding and short-term temperature stress on the content and isotopic signature of fatty acids,sterols, and alcohols in the scleractinian coral <Emphasis Type="Italic">Turbinaria reniformis</Emphasis>

This study assesses the combined effect of feeding and short-term thermal stress on various physiological parameters and on the fatty acid, sterol, and alcohol composition of the scleractinian coral Turbinaria reniformis. The compound-specific carbon isotope composition of the lipids was also measured. Under control conditions (26°C), feeding with Artemia salina significantly increased the symbiont density and chlorophyll content and the growth rates of the corals. It also doubled the concentrations of almost all fatty acid (FA) compounds and increased the n-alcohol and sterol contents. δ¹³C results showed that the feeding enhancement of FA concentrations occurred either via a direct pathway, for one of the major polyunsaturated fatty acid (PUFA) compounds of the food (18:3n-3 FA), or via an enhancement of photosynthate transfer (indirect pathway), for the other coral FAs. Cholesterol (C₂₇Δ⁵) was also directly acquired from the food. Thermal stress (31°C) affected corals, but differently according to their feeding status. Chlorophyll, protein content, and maximal photosynthetic efficiency of photosystem II (PSII) decreased to a greater extent in starved corals. In such corals, FA concentrations were reduced by 33%, (especially C16, C18 FAs, and n-3 PUFA) and the sterol content by 27% (especially the C₂₈∆^5,22 and C₂₈∆⁵). The enrichment in the δ¹³C signature of the storage and structural FAs suggests that they were the main compounds respired during the stress to maintain the coral metabolism. Thermal stress had less effect on the lipid concentrations of fed corals, as only FA levels were reduced by 13%, with no major changes in their isotope carbon signatures. In conclusion, feeding plays an essential role in sustaining T. reniformis metabolism during the thermal stress.     

Effects of starvation on moult cycle and hepatopancreas of Stage I lobster(Homarus americanus) larvae

Effects of feeding and starvation on the moult cycle and on the ultrastructure of hepatopancreas cells were studied in Stage I lobster larvae (Homarus americanus Milne-Edwards). The relative significance of yolk and first food was quite different in larvae originating from two females. This difference was evident also in the amounts of stored lipid in the R-cells of the larval hepatopancreas. Most larvae from one hatch were, in principle, able to develop exclusively with yolk reserves (without food) to the second instar. The larvae from the second hatch showed lecithotrophic development only to the transition between late intermoult and early premoult (Stages C/D₀ of Drachs's moult cycle) of the first larval instar. When initial starvation in this group lasted for 3 days or more, the point of no return (PNR) was exceeded. After the PNR, consumption of food was still possible, but development ceased in the transition C/D₀ or in late premoult (D_3–4). It is suggested that these stages of the moult cycle are critical points were cessation of development and increased mortality are particularly likely in early larval lobsters under nutritional stress. Examination of hepatopancreas R-cells suggested that the PNR is caused by an irreversible loss of the ability to restore lipid reserves depleted during initial starvation. Initial periods of starvation ending before the PNR prolonged mainly Stage D₀ of the same instar (I). During this delay, structural changes in the R-cells caused by the preceding period of starvation were reversed: reduced lipid inclusions, swollen mitochondria, an increased number of residual bodies indicating autolysis, and a reduction of the microvillous processes. Continually starved larvae which showed lecithotrophic development throughout the first instar and were then re-fed after moulting successfully, had later a prolonged intermoult (Stage C) period in the second instar. This shows that, despite occasional lecithotrophy, food is an important factor in early larval development of the lobster.     

Fatty acid composition of membrane bilayers: Importance of diet polyunsaturated fat balance

In one of the most extensive analyses to date we show that the balance of diet n-3 and n-6 polyunsaturated fatty acids (PUFA) is the most important determinant of membrane composition in the rat under 'normal' conditions. Young adult male Sprague-Dawley rats were fed one of twelve moderate-fat diets (25% of total energy) for 8weeks. Diets differed only in fatty acid (FA) profiles, with saturate (SFA) content ranging 8-88% of total FAs, monounsaturate (MUFA) 6-65%, total PUFA 4-81%, n-6 PUFA 3-70% and n-3 PUFA 1-70%. Diet PUFA included only essential FAs 18:2n-6 and 18:3n-3. Balance between n-3 and n-6 PUFA is defined as the PUFA balance (n-3 PUFA as % of total PUFA) and ranged 1-86% in the diets. FA composition was measured for brain, heart, liver, skeletal muscle, erythrocytes and plasma phospholipids, as well as adipose tissue and plasma triglycerides. The conformer-regulator model was used (slope=1 indicates membrane composition completely conforming to diet). Extensive changes in diet SFA, MUFA and PUFA had minimal effect on membranes (average slopes 0.01, 0.07, 0.07 respectively), but considerable influence on adipose tissue and plasma triglycerides (average slopes 0.27, 0.53, 0.47 respectively). Diet balance between n-3 and n-6 PUFA had a biphasic influence on membrane composition. When n-3 PUFA<10% of total PUFA, membrane composition completely conformed to diet (average slope 0.95), while diet PUFA balance>10% had little influence (average slope 0.19). The modern human diet has an average PUFA balance ~10% and this will likely have significant health implications.     

Starvation reduces the heat shock protein responses in white sturgeon larvae

This study investigates the responses of white sturgeon larvae (Acipenser transmontanus) to starvation and thermal stress, through the measurement of nutritional status (i.e. growth performances) and cellular biomarkers: heat shock proteins (Hsp) 70 and 90. White sturgeon larvae (25 day post hatch; initial weight 179.0 ± 5.1 mg) were fed (20% body weight per day) or starved for 24, 48 or 72 hrs. Every 24 hrs, five larvae from each of the starved or fed treatment replicates were exposed to heat shock resulting from an increase in water temperature from 19°C to 26°C, at a rate of 1°C per 15 min, and maintained at 26°C for 4 hrs. No mortality was observed in this study. Starvation significantly (p < 0.05) decreased the body weight and body contents of energy, protein, and lipid of the experimental larvae, compared to the fed larvae. Heat shock induced the expressions of Hsp70 and Hsp90 in both the fed and starved group; however, starvation reduced the induction at all sampling points. The current study demonstrates that poor larval nutritional status, assessed by the aforementioned parameters, reduced heat shock responses to thermal stress, as measured by heat shock protein levels. Furthermore, Hsp70 and 90 are more sensitive to heat shock and starvation, respectively. This may be, in part, a result of the different functioning of the heat shock proteins in cellular stress response and warrants further study.     

Dietary fatty acid composition changes mitochondrial phospholipids and oxidative capacities in rainbow trout red muscle

Dietary conditioning of juvenile trout changed the acyl chain composition of mitochondrial phospholipids and the oxidative capacities of muscle mitochondria. Trout were fed three diets differing only in fatty acid (FA) composition. The highly unsaturated 22:6 n-3 (DHA) accounted for 0.4, 14, and 30% of fatty acids in Diets 1, 2 and 3. After 10 weeks of growth, the dietary groups differed markedly in FA composition of mitochondrial phospholipids, with significant dietary effects for virtually all FA. Mean mitochondrial DHA levels were 19, 40 and 33% in trout fed Diets 1, 2 and 3. Mitochondrial oxidative capacities changed with diet, while mitochondrial concentrations of cytochromes and of the adenylate nucleotide translocase (nmol mg¹ protein) did not. Mitochondria from fish fed Diet 1 had higher non-phosphorylating (state 4) rates at 5°C than those fed other diets. When phosphorylating (state 3) rates differed between dietary groups, rates at 5 and 15°C were higher for fish fed the more unsaturated diets. Stepwise multiple regressions indicated that FA composition could explain much (42–70%) of the variability of state 4 rates, particularly at 5°C. At 15°C, FA composition explained 16–42% of the variability of states 3 and 4 rates. Similar conclusions were obtained for the complete data set (trout fed diets 1, 2 and 3) and for the data from trout achieving similar growth rates (e.g. those fed Diets 1 and 2). Neither general characteristics of membrane FA, such as % saturates, unsaturation index, n-3, n-6 or n-3/n-6 nor levels of abundant unsaturated FA such as DHA or 18:1(n-9 + n-7), were systematically correlated with mitochondrial capacities even though they differed considerably between trout fed the different diets. Relatively minor FA (20:5n-3, 20:0, 18:2n-6, 18:3n-3, 18:0 and 15:0) showed better correlations with mitochondrial oxidative capacities. This supports the concept that acyl chain composition modulates mitochondrial capacities via interactions between membrane proteins and specific FA of particular phospholipid classes in their microenvironment.     

Effects of starvation on body composition and cold tolerance in the collembolan Orchesella cincta and the isopod Porcellio scaber

The effects of long-term starvation on the body composition of the isopod Porcellio scaber (Latreille) and the collembolan Orchesella cincta (L.) were studied, by determining the body composition in starved and fed animals. A period under summer conditions (19 degrees C, 75% RH and L/D 16/8 photoperiod), was followed by a period under winter conditions (5 degrees C, 75% RH and LD 6/18 photoperiod). O. cincta was held under summer conditions for 3weeks, during which its protein and lipid content decreased, while its water content increased. In P. scaber, the same occurred during the 6weeks they were kept under summer conditions. During subsequent weeks under winter conditions, changes in cold tolerance of the animals were investigated. Cold tolerance and haemolymph osmolality were measured once a week. Starved animals had lower cold tolerance than fed ones. For P. scaber a decreased haemolymph osmolality was found in starved animals compared to fed ones. This is assumed to be caused by a combination of the consumption of carbohydrates out of the haemolymph and of protein reserves and the accumulation of body water. O. cincta appeared to be capable of osmoregulation, as haemolymph osmolality did not differ between starved and fed animals, despite differences in body water content. Decreased cold tolerance in starved animals of both species may be caused by increased water content or, more probably, by the decrease in reserves needed to produce cryoprotective substances.     

Dietary neutral lipid level and source in Senegalese sole (Solea senegalensis) larvae: effect on growth, lipid metabolism and digestive capacity

Contrary to larval essential fatty acid (EFA) requirements, the effect of dietary neutral lipid supply has been little investigated in marine fish larvae. The present work investigates the effect of feeding Senegalese sole larvae on Artemia enriched with higher or lower doses of lipid emulsion. Two lipid sources - soybean oil and fish oil - were compared. From 16 days after hatching (DAH) onwards, larvae were fed one of four experimental treatments: Artemia enriched on a high or low dose of soybean oil emulsion (HS and LS) or Artemia enriched on a high or low dose of fish oil emulsion (HF and LF). In terms of growth, the dietary lipid level did not have a significant effect while the soybean oil treatments induced a lower growth than the fish oil-enriched Artemia. The fatty acid (FA) composition of the larvae closely reflected the dietary quantitative and qualitative FA profile. Only slight dietary effects were noted in the activity of trypsin, lipase and alkaline phosphatase. A higher amount of lipid droplets was noticeable in the posterior intestine epithelia and in the hepatocytes of larvae fed Artemia enriched with higher lipid doses, while LS-Artemia induced the lower lipid accumulation on the basal zone of the enterocytes, in accordance with the lowest total lipid level measured in this treatment. These results suggest an important effect of dietary total lipid level on lipid accumulation in the enterocytes and on FA absorption. At 33 DAH a tube feeding trial was conducted with 14C-labelled oleic acid (OA) or triolein (TRI), showing that the lower accumulation of lipid droplets in the larvae fed LS was associated with a significantly higher absorption and retention in the gut and body tissues of the TRI label. For OA no significant differences between treatments were found. TRI label was considerably more evacuated than OA, indicating that sole larvae may have a lower capacity to incorporate a triacylglycerol, which needs to be digested. Finally, OA appears to be preferentially utilized for energy production, accumulating more in larval tissues when absorbed in higher amounts.     

Reproduction response of Chinese mitten-handed crab (Eriocheir sinensis) fed different sources of dietary lipid

The effect of feeding three semi-purified diets containing different lipid sources (anchovy oil, soybean oil and pork lard) on fecundity, hatchability and egg fatty acid composition of Chinese mitten-handed crab (Eriocheir sinensis) broodstock was compared with a fresh clam diet in a 6-month feeding trial. Broodstock crabs fed the diet containing pork lard showed poor fecundity and low hatchability. Crabs fed the diet containing soybean oil showed improved fecundity; however, no significant improvement in hatchability was observed. Broodstock fed the diet containing anchovy oil showed the highest fecundity and egg hatchability. Eggs from broodstock fed anchovy oil as sole dietary lipid had a higher n-3 polyunsaturated fatty acid (PUFA) content (33.3%) compared with those of crabs fed diets with soybean oil (20.1%) and pork lard (16.3%) as lipid sources. The results indicate a close correlation between: (1) the 20:5n-3 content of the egg lipid and fecundity; (2) the 22:6n-3 content and hatchability; and (3) fecundity, hatchability and n-3/n-6 fatty acid ratio. The results also suggest that each of these n-3 HUFAs may play different and specific roles in crab reproduction and that either must be adequate in the broodstock diet.     

Behavioural differences in starving herring Clupea harengus L. larvae correlate with body levels of essential fatty acids

Twenty days after hatching, a single stock of Atlantic herring ( Clupea harengus L.) larvae, cultured in the presence of rotifers and Artemia nauplii but showing little or no active feeding behaviour, displayed clear signs of starvation. Three groups of fish were distinguished: group I was generally pinhead-shaped, tended to swim with a spinning motion and floated vertically; group 2 lay moribund on the bottom of the tank; group 3 showed normal, active swimming behaviour. Fatty acids of total lipid extracted from groups 1 and 2 differed from group 3 in having markedly reduced percentages of 20:5n-3, 22:6n-3, 20:1 and 22:1. We conclude that individuals within a single stock of cultured herring larvae respond differently to starvation and that this generates well defined behavioural differences which correlate with levels of n-3 polyunsaturated fatty acids (PUFA) in body lipid. The implications of these observations are discussed.