A determination of the quantitative requirements for docosahexaenoic acid for juvenile barramundi (Lates calcarifer)

A series of diets with varying docosahexaenoic acid (DHA; 22:6n‐3) inclusion levels (1 g kg^?1 3 g kg^?1, 6 g kg^?1, 10 g kg^?1, 15 g kg^?1 and 18 g kg^?1) were fed to juvenile barramundi (Lates calcarifer) for 6 weeks. Two additional diets examined the addition of arachidonic acid (ARA; 20:4n‐6) or eicosapentaenoic acid (EPA; 20:5n‐3) to the diets at 10 g kg^?1 when DHA was also included at 10 g kg^?1. Fish were fed the diets on a pair‐fed feeding regime to eliminate feed intake variability. Fish were weighed, and blood and tissue samples were collected after 6 weeks. Behavioural parameters were also assessed. Improvement in growth was seen with increasing inclusion of DHA up to a maximum at 10 g kg^?1 inclusion, albeit the response was minor. However, the addition of ARA to the diet reduced the growth response, while the addition of EPA improved the growth response. An improvement in feeding behaviour was also seen with increasing DHA up to a peak at 10 g kg^?1, while those animals fed diets low in DHA showed increasingly cryptic behaviour. With the increasing inclusion of DHA, a range of pathologies were observed, but the addition of an EPA component to the diet limited these pathologies, while the addition of ARA made little improvement and in some cases exacerbated the pathologies.     

Effects of dietary docosahexaenoic acid sources,microalgae meal and oil,on growth,fatty acid composition and docosahexaenoic acid retention of orange‐spotted grouper,Epinephelus coioides

This study evaluated the effects of Aurantiochytrium spp. microalgae meal and oil as dietary docosahexaenoic acid (DHA) sources on the growth, fatty acid composition and DHA retention of orange‐spotted grouper, Epinephelus coioides. Dietary fish oil was replaced with microalgae meal or oil to provide an equal amount of DHA as a fish oil‐containing basal diet. In total, three experimental diets were fed to triplicate groups of fish (initial wt: 8.48 ± 0.06 g) in a recirculating system for 8 weeks. The weight gain and feed efficiency of the fish did not differ significantly among the experimental diets. The fatty acid composition of the whole body of the fish generally reflected the composition of their diet. The concentration of eicosapentaenoic acid in the whole body was higher in the fish fed the fish meal control diet than in those fed the two experimental diets The fish fed the control diet and those fed the diet containing microalgae oil exhibited higher DHA concentrations than did the fish fed the diet containing microalgae meal. The whole‐body DHA retention was the highest in the fish fed the diet with microalgae oil, followed by the fish fed the control diet. The lowest whole‐body DHA retention was observed in the fish fed the diet containing microalgae meal. The results suggested that the oil from Aurantiochytrium spp. microalgae can be used as DHA source for the grouper. DHA utilization by the fish was higher when the diet was supplemented with microalgae oil than with dry microalgae meal.     

Effects of different dietary arachidonic acid : docosahexaenoic acid ratios on phospholipid fatty acid compositions and prostaglandin production in juvenile turbot (Scophthalmus maximus)

Five purified diets containing AA (20:4n-6) at 0.02–0.78% dry weight and DHA (22:6n-3) at 0.93–0.17% dry weight were fed to duplicate groups of juvenile turbot (Scophthalmus maximus) of initial weight 0.87 g for a period of 11 weeks. The dietary DHA:AA ratio ranged from 62 to 0.2. Incorporation of AA into liver phospholipids increased with increasing dietary AA input. Phospholipids from fish fed diets containing 0.02, 0.06 and 0.11% of dry weight as AA generally contained less AA compared to fish fed fish oil while those fed diets containing 0.35 and 0.78% of dry weight as AA had higher AA levels in their phospholipids. The highest levels of AA were found in PI but the greatest percentage increase in AA incorporation was in PE and PC. Brain phospholipid fatty acid compositions were less altered by dietary treatment than those of liver but DHA content of PC and PE in brain was substantially lower in fish fed 0.93% pure DHA compared to those fed fish oil. This suggests that dietary DHA must exceed 1% of dry weight to satisfy the requirements of the developing neural system in juvenile turbot. In both tissues, (20:5n-3) concentration was inversely related to both dietary and tissue PI AA concentration. Similar dietary induced changes in AA, EPA and DHA concentrations occurred in the phospholipids of heart, gill and kidney. PGE₂ and 6-ketoPGF₁ were measured in homogenates of heart, brain, gill and kidney. In general, fish fed the lowest dietary AA levels had reduced levels of prostaglandins in their tissue homogenates while those fed the highest level of AA had increased prostaglandin levels, compared to fish fed fish oil. In brains, the PGE₂ concentration was only significantly increased in fish fed the highest dietary AA.Abbreviations AA arachidonic acid - DHA docosahexaenoic acid - EFA essential fatty acid - EPA eicosapentaenoic acid - HPTLC high performance thin-layer chromatography - HUFA highly unsaturated fatty acid - PC phosphatidylcholine - PE phosphatidylethanolamine - PGE prostaglandin E - PGE prostaglandin E - PI phosphatidylinositol - PS phosphatidylserine - PUFA polyunsaturated fatty acid - TLC thin-layer chromatography     

No significant effect of additive ratios of docosahexaenoic acid to eicosapentaenoic acid on the survival and growth of cobia (Rachycentron canadum) juvenile

An experiment was conducted in the laboratory to investigate the effects of additive ratios of docosahexaenoic acid (DHA) to eicosapentaenoic acid (EPA) on the growth and survival of cobia (Rachycentron canadum) juveniles from August to October 2005. Three hundred and eighty cobia juveniles (56 days of age, body weight 6.9 ± 0.1 g, body length 9.2 ± 0.1 cm) were selected and 20 of them were freely taken for initial sample analysis in the week 0. Additional 360 juveniles were randomly assigned into eight groups with triplicate, total 24 tanks with 15 fish each. Cobia juveniles were reared in glass‐steel tanks (200‐L volume per tank) using filtered seawater with temperature 26–30.5 °C, salinity 25.4–33.0 g L^?1 and pH 7.8–8.0. Cobia juveniles were fed for 8 weeks using seven treatment diets (D‐1 to D‐7) with the same amount of DHA and EPA (15.0 ± 1.2 g kg^?1 of dried diet), but varying ratios of DHA to EPA (0.9, 1.1, 1.3, 1.5, 1.7, 1.9, 2.1, respectively) and a control diet (D‐0, DHA + EPA = 8.0 g kg^?1 of dried diet, DHA/EPA = 1.3). Five juveniles per tank were randomly taken for sample analysis at the end of weeks 4 and 8, respectively. The highest protein efficiency rate (PER; 1.5 in mean), average body weight (BW; 73.3 g per fish in mean) and the lowest feed conversion ratio (FCR; 1.6 in mean) were obtained in cobia juveniles fed the control diet at the end of week 8. These parameters were significantly different (P < 0.05) among juveniles fed the control and treatment diets; however, no significant difference (P > 0.05) was found among juveniles fed the treatment diets evaluated in this study. It was concluded that the survival and growth of cobia juveniles were not greatly influenced by additive ratios of DHA to EPA in our experimental conditions.     

Lipid class and fatty acid composition of phospholipids from the gonads of skipjack tuna

ABSTRACT:   Lipid class and fatty acid composition of phospholipids from the gonads of skipjack tuna were examined to evaluate effective utilization of the processing of by-products. The predominant phospholipids in the ovaries were phosphatidylcholine (PC; 47.9%), phosphatidylethanolamine (PE; 19.3%) and lyso-phosphatidylcholine (LPC; 19.1%). In contrast, those in the testes were PC (40.1%), PE (29.3%) and phosphatidylserine (PS; 9.6%). The percentage of docosahexaenoic acid (DHA) was markedly high at more than 50% in LPC of the ovaries, and PE and PS of the testes were also high. The percentages of DHA at sn-position 2 of the predominant phospholipids, except for PC in the testes, were more than 60%, in particular PE in the testes was remarkably high at 81.9%. After storage for 2 days at 5°C, the LPC content in the ovaries increased twofold and the DHA level of LPC was the same as before, though the contents of other phospholipids decreased.     

Time kinetics of fatty acid changes in phospholipids following enrichment and starvation of Artemia franciscana with a main focus on docosahexaenoic acid

The main objective was to study time kinetics of change in important highly unsaturated fatty acids (HUFAs) in phosphatidylcholine (PC) and phosphatidylethanolamine (PE) of Artemia franciscana nauplii and juveniles following enrichment and subsequent starvation. Samples of Artemia nauplii were taken at variable times (0.5–24 h) following enrichment and starvation. Samples of Artemia juveniles were taken after 2, 3 and 4 days of cultivation. No docosahexaenoic acid (DHA) was found in PC and PE of Artemia nauplii during the first hour of enrichment, while a significant (P < 0.05) increase was found in total lipids (TLs). The content of DHA in PC and PE increased thereafter steadily from 1 to 8 h of enrichment. DHA in PC and PE during enrichment (1–8 h) and following starvation (8–24 h), respectively, increased and decreased significantly (P < 0.05), but at a lower rate than that in TL. Moreover, juvenile Artemia (2–4 days) contained a relatively low level of DHA in TL compared with enriched Artemia nauplii, but the content of DHA in PC and PE was similar. The results open perspectives for both industry and science. For scientific studies, the lag phase in HUFA enrichment makes it possible to produce Artemia nauplii with variable relative HUFA enrichments in phospholipids and TL.     

Changes in proximate composition,lipid class and fatty acid profile in Greenshell™ mussels (Perna canaliculus) over an annual cycle

Greenshell? mussel (GSM, Perna canaliculus) is New Zealand's most important aquaculture species. This study looked at changes in the proximate composition, lipid class and fatty acid profile of male and female market ready mussels over a year from April 2016 through to March 2017. There were differences over the season as well as differences between male and female mussels. Winter GSM had lower lipid, carbohydrate and protein concentrations than other seasons. Female mussels had a higher lipid (on average 1.8 ± 0.5 g/100 g ww) and carbohydrate (on average 5.0 ± 1.1 g/100 g ww) compared with the males (lipid, 1.5 ± 0.4 g/100 g ww and carbohydrate 4.5 ± 1.2 g/100 g ww). The major differences observed between seasons in the omega 3 and fatty acids content of the GSM were driven by the fat content, with the highest fat content found in female mussels in the spring prior to spawning. The majority of lipids found in mussels are polar lipids. Overall, this study provides the most comprehensive data on the composition and the lipid content of GSM to date and provides solid base‐line data as the New Zealand GSM industry continues to improve their production procedures. The enhanced nutritional payload of mussels harvested outside of winter (particularly female mussels in spring) could drive a differentiated higher value mussel product.     

Fatty acid composition,osmolality, Na+, K+‐ATPase activity,cortisol content and antioxidant status of rainbow trout (Oncorhynchus mykiss) in response to various dietary levels of eicosapentaenoic acid and docosahexaenoic acid

This study was conducted to evaluate the effect of dietary eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) levels on the fatty acid composition, salinity tolerance and antioxidant status of rainbow trout (Oncorhynchus mykiss). Four diets were formulated with total EPA and DHA contents of 5.41, 9.55, 13.97 and 17.88 g/kg (abbreviated as ED‐5.41, ED‐9.55, ED‐13.97 and ED‐17.88 respectively). Rainbow trout (initial weight of 90.61 ± 9.25 g) were fed the experimental diets for 8 weeks to accumulate significant differences in fatty acid composition and subsequently underwent salinity acclimation. Our results indicated that high dietary EPA and DHA significantly improved the EPA and DHA content in fish tissues. The serum osmolality of fish returned to their freshwater values in the ED‐9.55, ED‐13.97 and ED‐17.88 groups. The Na⁺, K⁺‐ATPase (NKA) activity of fish in the ED‐13.97 group changed dramatically to adapt the fish to the hypertonic environment. Moreover, there was no significant difference in the serum cortisol concentration and liver catalase (CAT) activity of fish in the ED‐13.97 group during salinity acclimation. The liver superoxide dismutase (SOD) activity in the ED‐13.97 group was significantly higher than that in ED‐5.41 and ED‐9.55 groups at the end of salinity acclimation. The muscle malondialdehyde (MDA) content in the ED‐13.97 group was significantly lower than that in the ED‐17.88 group before salinity acclimation and significantly lower than the ED‐5.41 and ED‐17.88 groups on day 7 of acclimation. The results of this study indicate that the rainbow trout in the ED‐13.97 group exhibited optimal salinity acclimation performance.     

Oxidative status and histological changes in sea bass larvae muscle in response to high dietary content of docosahexaenoic acid DHA

In previous studies, we observed dystrophic alterations in muscle of 48‐day‐old sea bass fed imbalanced docosahexaenoic acid (DHA) and vitamin E diets. To understand the whole pathological process associated with oxidative stress, a histological study was performed by feeding 14‐day‐old sea bass larvae with microdiets containing different ratios of DHA/vitamin E (1/150, 5/150 and 5/300) for a period of 21 days. Larvae fed diet 1/150 showed no lesions in contrast to larvae fed diets 5/150 and 5/300 where the highest incidence of muscle lesions and thiobarbituric acid reactive substances (TBARS) content was observed. Semithin sections revealed focal lesions consisting of degenerated fibres with hypercontracted myofilaments and extensive sarcoplasm vacuolization affecting both red and white muscle. Ultrathin sections of degenerating muscle fibres showed diffuse dilatation of sarcoplasmic reticulum, disorganized myofilaments and autophagic vacuoles containing myelin figures and dense bodies. Additionally, some macrophages were observed among injured fibres as numerous satellite cells. Results from the study agree with those obtained from previous work, proving the pathological potential of free radicals in sea bass larvae musculature. Moreover, high vitamin E inclusion could not completely protect cell membranes from free radicals action.     

Effect of increasing docosahexaenoic acid content in weaning diets on survival,growth and skeletal anomalies of longfin yellowtail (Seriola rivoliana,Valenciennes 1833)

Five isoproteic (54.8%) and isolipidic (24.1%) microdiets, which varied in their docosahexaenoic acid (DHA) content (0.25%, 0.75%, 1.64%, 1.99% and 3.17%; dw), were manufactured to determine its effects on longfin yellowtail Seriola rivoliana larvae in terms of fish biological performance, whole body fatty acid profile and incidence of skeletal anomalies from 30 dah (11.31 ± 1.79 Total Length, TL) to 50 dah (19.80 ± 0.58 mm TL). The inclusion of dietary DHA up to 3.17% (dw) improved larval resistance to air exposure, although DHA did not significantly affect fish final growth or final survival. Indeed, high levels of dietary DHA (1.99% and 3.17%, dw) tended to increase the incidence of skeletal anomalies in S. rivoliana larvae, albeit no significant differences were observed. Furthermore, the occurrence of severe anomalies such as kyphosis and lordosis, was mainly associated to the larvae fed the highest levels of dietary DHA. In terms of survival, increasing dietary DHA levels did not significantly affect longfin yellowtail survival rate, despite a tendency for enhanced survival. The results of the present study proved that the inclusion of dietary DHA in inert diets up to a 3.17% (dw) and a DHA/EPA ratio above 3.1 increased the final survival and stress resistance in S. rivoliana larvae.     

模拟探究凡纳滨对虾磷脂在贮藏过程中的水解机理

为探究凡纳滨对虾磷脂(PL)在贮藏过程中的水解机理,本研究建立了磷脂相关酶与磷脂酰胆碱(PC)的体外模型,模拟凡纳滨对虾磷脂水解反应。实验分别从凡纳滨对虾体内分离纯化得到磷脂相关酶与PC,进行体外反应,并采用"鸟枪"脂质组学法及气相色谱—质谱法分析水解反应前后PC、溶血磷脂(LPL)及游离脂肪酸(FFA)的组成和含量变化,进而推断磷脂的水解机理。结果显示,凡纳滨对虾体内磷脂水解酶包括脂肪酶、磷脂酶A_1(PLA_1)、磷脂酶A_2(PLA_2)、磷脂酶C(PLC)及磷脂酶D(PLD),其中PLA2活力最高,可达177.87 U。体外模拟反应中,凡纳滨对虾PC含量由516.45显著降至146.14 mg/g,总FFA含量由36.42上升至568.57 mg/g,其中多不饱和脂肪酸(PUFA)显著增加了280.5 mg/g。通过相关性分析水解前后磷脂水解酶活力变化与PC含量变化,发现PC的变化与PLA2显著相关(R=0.91)。研究表明水产品中PC变化是磷脂相关酶催化的水解反应,其水解产物主要为FFA与LPC,且PLA_2对水产品贮藏过程中磷脂的水解影响最大。本实验建立了新型的体外水解模型,并初步探究了磷脂的水解机理,为水产品贮藏及磷脂酶的研究及应用奠定了理论基础。     

Organ and phospholipid class fatty acid specificity in response to dietary depletion of essential n‐3 fatty acids in Atlantic salmon (Salmo salar L.)

The cell membrane phospholipid composition is of major importance for normal cell functions. However, it is not known how complete depletion of both shorter and longer chain omega‐3 fatty acids in salmon diets influences fatty acid composition of phospholipid subclasses in different organs of Atlantic salmon. We describe here the fatty acid composition in phospholipid subclasses of liver, muscle, heart and intestine in Atlantic salmon after 18 months of dietary n‐3 essential fatty acids deprivation. The percentage of 22:6n‐3 was markedly reduced in almost all phospholipid subclasses, and except for muscle phosphatidylcholine, phosphatidylethanolamine (PE) and phosphatidylinositol (PI), phospholipids in deficient fish were totally devoid of 20:5n‐3. As compensation, we observed significant increases in 20:4n‐6, and especially in 20:3n‐9 (Mead acid) and 22:5n‐6, varying among phospholipids and organs. High amounts of 20:3n‐9 were found in liver and intestinal PE, little in PE from heart and muscle. For 22:5n‐6, we saw a small incorporation in PI in liver and intestine compared to heart and muscle. Generally in PI, the preference for 20:4n‐6 to 20:5n‐3 differed significantly between organs. Overall, changes upon n‐3 deprivation seemed to be strongest in liver and intestine, the lipid‐secreting organs, and less in muscle and heart.     

Effects of dietary docosahexaenoic acid on growth performance,fatty acid profile and lipogenesis of blunt snout bream (Megalobrama amblycephala)

Six diets were designed to investigate the effects of dietary docosahexaenoic acid (22:6n‐3; DHA) levels (0.5, 1.3, 2.3, 4.2, 8.1 and 15.9 g/kg diets) on growth performance, fatty acid profile and expression of some lipogenesis‐related genes of blunt snout bream (Megalobrama amblycephala). Fish (average weight: 26.40 ± 0.11 g) were randomly fed one of six diets for 8 weeks. Results indicated that the final body weight (FBW) and specific growth rate (SGR) of fish fed 1.3 g/kg DHA were significantly higher than other groups except for the 2.3 g/kg DHA (p < .05). Compared with other groups, the number of lipid droplet clusters of the liver stained with oil red O in the 2.3 g/kg DHA group was the highest, which was consistent with the lipid contents of whole body and liver. The DHA proportion in liver and muscle significantly increased with the increasing dietary DHA levels (p < .05), which reflected fatty acid profiles of diets. The highest mRNA expressions of acetyl‐CoA carboxylase α (ACCα), fatty acid synthase (FAS) and sterol regulatory element‐binding protein‐1 (SREBP‐1) occurred in the 1.3 g/kg DHA group, followed by 2.3 g/kg DHA. In summary, the supplementation of 1.3–2.3 g/kg DHA could improve growth performance and lipogenesis, and the dietary DHA could improve DHA and PUFA proportion in liver and muscle.     

Effects of Artemia enriched with eicosapentaenoic and docosahexaenoic acid on survival and occurrence of molting failure in megalop larvae of the mud crab Scylla serrata

ABSTRACT:   The present study was conducted to investigate the effect of eicosapentaenoic acid (20:5n-3, EPA) and docosahexaenoic acid (22:6n-3, DHA) on the survival and the occurrence of molting failure to megalops of mud crab Scylla serrata larvae fed enriched Artemia . Survival rate, intermolt period, carapace width at the first crab stage, ratio of chela to carapace length at the fifth zoeal stage and the occurrence of molting failure to megalops were observed. Mud crab larvae were reared in 1-L plastic beakers and fed with Artemia enriched at five different levels of EPA (0.31% to 1.36% EPA; referred to as E25, E50, E75 and E100) or four different levels of DHA (0.75–0.95% EPA and 0.49–1.38% DHA; referred to as D25, D50, D75 and D100). As a negative control, larvae were fed Artemia enriched with oleic acid (OA). Mud crab larvae fed Artemia containing low (0.41–0.45% EPA and trace DHA) or high (1.36% and 0.95% EPA and 0.16% and 1.38% DHA) amounts of EPA and DHA showed a significantly lower survival rate and prolonged intermolt period ( P  < 0.05). Moreover, a high frequency of molting failure to the megalops stage (34 and 33%) occurs coincident with a high chela to carapace length ratio (43 and 44%) in mud crab larvae fed high amounts of EPA and DHA (E100 and D100), suggesting that both of these treatments contain EPA and DHA in excess. These results indicate that during Artemia feeding, EPA and DHA content should be adjusted to 0.71–0.87% and 0.49–0.72% for maintaining a high survival, accelerating the intermolt period, and producing larger carapace width in the first crab stage.     

Dietary docosahexaenoic acid to eicosapentaenoic acid (DHA/EPA) ratio influenced growth performance,immune response,stress resistance and tissue fatty acid composition of juvenile Japanese seabass,Lateolabrax japonicus (Cuvier)

A 10‐week feeding experiment was conducted to investigate the effects of dietary DHA/EPA ratio on juvenile Japanese seabass reared in sea floating cages. Six practical diets were formulated differing only in DHA/EPA ratio: 0.55 (Diet D/E0.55), 1.04 (D/E1.04), 1.53 (D/E1.53), 2.08 (D/E2.08), 2.44 (D/E2.44) and 2.93 (D/E2.93). All diets had the same contents of total n‐3 long‐chain polyunsaturated fatty acid (LC‐PUFA) and arachidonic acid (ARA). The results showed that the final weight and specific growth rate significantly increased with increasing dietary DHA/EPA ratio from 0.55 to 2.08 and thereafter declined. Activities of lysozyme and superoxide dismutase in serum in groups with DHA/EPA of 1.53–2.93 was significantly higher compared to group D/E0.55 while the activity of serum alternative complement pathway in group D/E2.93 was significantly lower compared with group D/E1.53. However, no difference was observed in activities of both respiratory burst of head kidney macrophage and serum catalase among dietary treatments. The per cent survival after air exposure in group D/E1.53, D/E2.08 and D/E2.93 was significantly higher compared with group D/E0.55. The fatty acid composition of whole body and tissues reflected closely those of diets, while fish accumulated more DHA than EPA in fish tissues, especially in muscle. These results suggested that at the same dietary contents of n‐3 LC‐PUFA (appr. 18% of TFA) and ARA (appr. 0.7% of TFA), moderate dietary DHA/EPA ratios of 1.53–2.08 significantly enhanced growth performances, certain innate immune responses, and the stress tolerance of Japanese seabass, in accordance with the preferential incorporation of DHA into fish tissues.