Development of a High-Performance Thin-Layer Chromatography Method for the Quantification of Alkyl Glycerolipids and Alkenyl Glycerolipids from Shark and Chimera Oils and Tissues

Ether lipids are composed of alkyl lipids with an ether bond at the sn-1 position of a glycerol backbone and alkenyl lipids, which possess a vinyl ether bond at the sn-1 position of the glycerol. These ether glycerolipids are present either as polar glycerophospholipids or neutral glycerolipids. Before studying the biological role of molecular species of ether glycerolipids, there is a need to separate and quantify total alkyl and alkenyl glycerolipids from biological samples in order to determine any variation depending on tissue or physiopathological conditions. Here, we detail the development of the first high-performance thin-layer chromatography method for the quantification of total alkyl and alkenyl glycerolipids thanks to the separation of their corresponding alkyl and alkenyl glycerols. This method starts with a reduction of all lipids after extraction, resulting in the reduction of neutral and polar ether glycerolipids into alkyl and alkenyl glycerols, followed by an appropriate purification and, finally, the linearly ascending development of alkyl and alkenyl glycerols on high-performance thin-layer chromatography plates, staining, carbonization and densitometric analysis. Calibration curves were obtained with commercial alkyl and alkenyl glycerol standards, enabling the quantification of alkyl and alkenyl glycerols in samples and thus directly obtaining the quantity of alkyl and alkenyl lipids present in the samples. Interestingly, we found a differential quantity of these lipids in shark liver oil compared to chimera. We quantified alkyl and alkenyl glycerolipids in periprostatic adipose tissues from human prostate cancer and showed the feasibility of this method in other biological matrices (muscle, tumor).     

Lipids in Wheat Flour Streams

Lipids play an important role in determining the quality of wheat flour and its suitability for different bakery products. The lipid content and fatty acid composition in various flour streams showed a wide variation. The free lipid content varied from 0·1 to 1·9% (db) and bound lipid content from 0·2 to 2·1% (db) in different flour streams. The amount of bound lipid was more in reduction streams. The palmitic and linoleic acids were the predominant fatty acids present in free and bound lipids. These fatty acids were found to be higher in the «III» «Break coarse», «IV» «Break filter», «III» «Break fine», «1st» and «4th» «middling». The studies showed that the reduction passage flours had more of bound lipids indicating the presence of polar lipids.     

Lipids,buoyancy and the seasonal vertical migration of Calanus finmarchicus

The copepod Calanus finmarchicus remains in diapause for up to 5 months in the cold (<0.5°C) deep (>700 m) waters of the Faroe–Shetland Channel of the north-western approaches to the North Sea. While in diapause, C. finmarchicus has a high lipid content, up to 76% of dry weight, mostly in the form of wax esters. The question we address here is how copepods with such a high content of buoyant lipids can remain in diapause at depth for an extended period of time? The corollary to this is how this lipid content hinders and/or assists the copepods in their seasonal vertical migration? Part of the answer is due to the physical properties of wax esters. These have a thermal expansion and compressibility higher than that of sea water. Thus, depending on their relative composition (i.e. wax esters/water/protein/chitin), a copepod that is positively buoyant in warm surface waters can become neutrally buoyant in cold deep water. We develop a simple three component physical model of a copepod to explore how and where they attain neutral buoyancy, how the lipid content can aid in their ascent, and what fraction of the lipids can be utilized in ascent in gonad/egg formation while maintaining observed ascent rates. As well as being an energy reserve, the results show that rather than being a barrier to vertical migration, wax esters serve as an important regulator of buoyancy.     

Recent advances in European sea bass and gilthead sea bream nutrition

European sea bass (Dicentrarchus labrax) and gilthead sea bream(Sparus aurata) are amongst the most important finfish speciescultured in the Mediterranean region. Production of these species isnowadays a well-controlled process, but knowledge of their nutritionalrequirements is still very limited. Nevertheless, a considerable amountof data has been accumulated in recent years, and the purpose of thispaper is to review the recent advances on the nutritional requirementsof sea bass and sea bream. The optimum protein to energy ratio of thediets of sea bass and sea bream seem to be higher than for salmonids,and there is some evidence that high dietary lipid levels have nobeneficial effects on fish performances. Although the essential aminoacid requirements were estimated by the ideal protein method, data basedon the dose-response method is only available for a few amino acids.Essential fatty acid requirements were estimated for sea bream juvenilesbut data is lacking for sea bass. Vitamin and mineral requirements ofthese species are practically unknown. Although the importance ofbroodstock nutrition on gonadal development, spawning and egg quality isrecognized, few studies were done to elucidate these aspects. The recentdevelopment of microparticulate diets for larvae will contribute to theaccurate evaluation of their nutritional requirements.     

Seasonal cycle of body composition and energy of brown trout (Salmo trutta) in a temperate zone lake

Abstract – Seasonal changes in protein and fat were investigated in brown trout ( Salmo trutta L.) of a stunted population from a small, temperate zone lake. The lake was ice-covered during winter (about 200 days), and hypolimnic water temperature during winter was about 4°C. During winter there was an increase in specific fat, in particular among the larger sized fish, while there was a general decrease in specific protein content among both small (<5 winters) and large (age 5 or older) trout. During winter (end of October to mid-May) an average trout gained 32 kJ of fat, but protein energy content decreased by 14 kJ, yielding a net energy increase of 18 kJ or a daily energy gain of 0.09 kJ  ·  day⁻¹. During the ice-free season (mid-May to the end of October) the trout increased mainly in protein content with daily energy gains of about 1.4 kJ  ·  day⁻¹, a value about 14 times higher than the corresponding winter value. Trout living in lakes may store considerable amounts of fat during wintertime in contrast to the depletion of fat reserves found among stream-living trout in the same area, which face lower water temperatures in the winter period. ^Note     

The effect of different levels of dietary phospholipid on growth, survival and nutrient composition of early juvenile cobia (Rachycentron canadum)

This experiment was conducted to evaluate the effect of different levels of dietary phospholipid (PL) on growth, survival and nutrient composition of 25 days posthatch cobia Rachycentron canadum (0.4 g initial wet weight). For 42 days, fish were fed fish meal and protein hydrolysate based diets containing four PL levels (0, 20, 40 and 80 g kg^?1dry matter: purity 97%) and phosphatidylcholine purity was 60%. All diets were isonitrogenous and isolipidic by regulating the fish oil and maize oil levels. Weight gain (2601–10892%), specific growth ratio (7.82–11.18) and survival (49–100) were significantly affected by dietary PL. Intraperitoneal fat ratio (0.19–0.74) and hepatosomatic index (2.67–3.08) increased with dietary PL level. The effect of dietary PL levels on the chemical composition of tissues was significant only for whole body and liver. The contents of plasma total cholesterol (2.47–3.77 mmol L^?1) and PL (1.03–2.97 mmol L^?1) increased with an increase in dietary PL. In conclusion, in our study survival and growth continued to increase even at the highest PL levels used (80 g kg^?1); therefore optimal dietary PL levels may well exceed 80 g kg^?1 for early juvenile cobia requirement. It also indicated from the experiment that PL could affect lipid deposition and resulted in a higher lipid level in fish tissue.     

大豆浓缩蛋白对中华鲟幼鱼生长、血脂和体成分的影响研究

分别用大豆浓缩蛋白(SPC)替代饲料中0(R0,对照组),15%(R15),30%(R30)和45%(R45)的鱼粉蛋白,配制成4组等氮等能饲料,喂养(58.00±2.69)g中华鲟(Acipenser sinensis)幼鱼10周,每组设3个重复,研究SPC对中华鲟幼鱼生长性能、饲料利用率,以及血脂和体成分的影响。结果显示,随替代比例的增加,中华鲟幼鱼的增重率(WG)、特定生长率(SGR)、饲料效率(FE)和蛋白质效率(PER)逐渐降低,肥满度和肝体指数无显著性差异(P>0.05)。FE在R30组出现显著差异(P<0.05),WG、SGR和PER在R45组出现显著性差异(P<0.05)。随替代比例的增加,血浆高密度脂蛋白胆固醇含量无显著变化(P>0.05),胆固醇(CHOL)、甘油三脂(TG)和低密度脂蛋白胆固醇(LDL-C)含量逐渐降低。CHOL、TG和LDL-C含量在R30组出现显著差异(P<0.05)。随替代比例的增加,肌肉和肝脏水分逐渐升高,脂肪逐渐降低,蛋白含量无显著差异(P>0.05)。R30和R45组的肌肉和肝脏水分含量显著高于R0组(P<0.05),脂肪含量显著低于R0组(P<0.05)。结果表明:在本试验条件下,综合考虑生长性能、饲料利用、血脂和体成分等指标,中华鲟幼鱼饲料中SPC替代鱼粉蛋白的比例可达30%。     

Effects of six alternative lipid sources on growth and tissue fatty acid composition in Japanese sea bass (Lateolabrax japonicus)

Ten-week experiment was carried out on Japanese sea bass (5.87 ± 0.02 g) to study the effects of replacement of fish oil with six alternative lipid sources: pork lard, PL; beef tallow, BT; poultry fat, PF; soybean oil, SO; corn oil, CO; and a mixed-fat (MF: tallow, 60%; soy oil, 20%; fish oil, 20%) on growth performance and fatty acid (FA) composition in fillet and liver. Seven isoenergetic and isonitrogenous experimental diets were formulated, containing 10% of added lipid. Fish oil was used in control diet, which was substituted by 50% with the alternative lipid sources in the other six diets.

Weight gain (WG), specific growth rate (SGR), Feed conversion ratio (FCR) feed intake and hepatosomatic index (HSI) of fish fed the experimental diets were not significantly different (P > 0.05). Protein efficiency ratio (PER) in fish fed the PF diet were significantly lower than those of fish fed SO and CO diets. Significant differences in carcass moisture and lipid contents of carcass and liver were observed among fish fed the dietary treatments. Generally, the fatty acid composition of fish fillets and livers reflected the dietary FA composition.     

Biomarkers of bone development and oxidative stress in gilthead sea bream larvae fed microdiets with several levels of polar lipids and α‐tocopherol

Although dietary marine phospholipids are able to improve culture performance of marine fish larvae in a further extend than soybean lecithin, both types of phospholipids (PL) markedly increase oxidative risk. The inclusion of a fat‐soluble antioxidant such as the vitamin E α‐tocopherol could allow a better control of oxidative stress. The objective of this study was to determine the combined effect of graded levels of α‐tocopherol with different levels and sources of krill phospholipids (KPL) and soybean lecithin (SBL) on growth, survival, resistance to stress, oxidative status, bone metabolism‐related genes expression and biochemical composition of sea bream larvae. Sea bream larvae were completely weaned at 16 dph and fed for 30 days seven microdiets with three different levels of PL (0, 40 and 80 g kg^?1 diet) and two of α‐tocopherol 1500 and 3000 mg kg^?1 diet. Sea bream larvae fed diets without PL supplementation showed the lowest survival, growth and stress resistance, whereas increase in PL, particularly KPL, markedly promoted larval survival and growth. However, feeding SBL markedly increased TBARs and GPX gene expression increasing the peroxidation risk in the larvae. Besides, KPL inclusion improved incorporation of n‐3 HUFA and, particularly, EPA into larval tissues, these fatty acids being positively correlated with the expression of BMP‐4, RUNX 2, ALP, OC and OP genes and to bone mineralization for a given larval size class. The increase in dietary α‐tocopherol tends to improve growth in relation to the n‐3 HUFA levels in the diet, denoting the protective role of this vitamin against oxidation. Indeed, dietary α‐tocopherol decreased the oxidative stress in the larvae as denoted by the reduction in larval TBARs contents and gene expression of SOD and CAT, but not GPX. Thus, increase in dietary α‐tocopherol effectively prevented the formation of free radicals from HUFA, particularly EPA, but did not affect the incidence of bone anomalies or the expression of genes related to osteogenetic processes.     

Effects of dietary fish oil replacement on growth and carcass proximate composition of juvenile barramundi (Lates calcarifer)

An experiment was carried out to determine the effects of dietary fish oil replacement at an optimal level on the growth and carcass proximate composition of juvenile barramundi or sea bass (Lates calcarifer). Ten fish feeds were formulated to contain iso‐ingredients with theoretically identical dietary lipid levels but with different sources of lipids (fish oil, soybean oil, canola oil and linseed oil) replacing dietary fish oil. Three hundred fish were equally divided into 30 70‐L plastic tanks (three replicate tanks for each treatment). Fish were fed with experimental diets for 40 days within closed recirculating freshwater systems (70 L h^?1, 28°C and 12L:12D). Growth, specific growth rate, food conversion ratio (FCR) and carcass proximate composition were determined. Dietary lipids significantly affected (P<0.05) the growth of the fish. Good growth and low FCR were observed in all treatments. Therefore, fish oils can be partially replaced by vegetable oils to reduce the feed cost. There were no significant differences (P>0.05) in lipid, moisture and protein content in fish carcasses among the group. However, juvenile barramundi showed increasing lipid and moisture content in muscle, whereas decreasing protein content when compared to the initial fish.