Effect of retinoic acid on gene expression profiles of bovine intramuscular preadipocytes during adipogenesis

To investigate genes involved in intramuscular adipogenesis in ruminants, 16 genes with dramatic variable expression were selected. These were selected from the differentiation‐ and proliferation‐phase libraries of our previous serial analysis of gene expression (SAGE) studies of a clonal bovine intramuscular preadipocyte (BIP) cell line. We harvested the BIP cells over 12 days after adipogenic stimulation with all‐trans retinoic acid (ATRA). Quantitative real‐time PCR confirmed the earlier SAGE study results of the expression patterns of 15 of the genes. On day 6, TG accumulation increased significantly in the BIP cells but was completely inhibited in the 3T3‐L1 cells (the monogastric reference). ATRA enhanced expression levels of six genes whereas it suppressed expression of eight genes on day 3 of adipogenesis in the BIP cells. Forty‐eight hours after transfection, the messenger RNA expression level of the adipose differentiation‐related protein (ADFP), encoded by one of the upregulated genes, in the ADFP small interference RNA (siRNA)‐transfected cells was 3.5% of that in negative control‐transfected cells. Also, 6 days after induction the TG level in the ADFP siRNA‐transfected cells was 21.8% lower than that in negative control‐transfected cells. This analysis of gene expression profiles after ATRA treatment will contribute to our understanding of the molecular mechanisms involved in bovine intramuscular adipogenesis.     

Effect of diacylglycerol acyltransferase 2overexpression in 3T3-L1 is associated to an increase in mono-unsaturated fatty acid accumulation

Background：Fatty acid（FA） composition is the most important parameter affecting the flavor and nutritional value of the meat.The final and the only committed step in the biosynthesis of triglycerides is catalyzed by diacylglycerol acyltransferase 2（DGAT2）.The role of DGAT2 in lipid accumulation has been demonstrated in adipocytes,However,little is known about the effect of DGAT2 on the FA composition of these cells.Methods：To investigate the role of DGAT2 in regulating lipid accumulation,FA composition and the expression of adipogenic genes,we cloned the open reading frame of the porcine DGAT2 gene and established 3T3-L1 cells that overexpressed DGAT2.Cells were then cultured in differentiation medium（DM） without FA,with a mixture of FAs（FA-DM）,or containing a ~（13）C stable isotope-labeled FA mixture（IFA-DM）.The FA composition of adipocytes was analyzed by gas chromatography-mass spectrometry and gas chromatography-isotope ratio mass spectrometry.Quantitative PCR and western blotting were employed to detect expression of adipogenic genes in 3T3-L1 adipocytes cultured with FA-DM for 12 d.Results：The triacylglyceride（TAG） content was significantly higher in 3T3-L1 adipocytes overexpressing DGAT2 than in control cells.When cultured in DM or FA-DM for 12 d,cells overexpressing DGAT2 showed a higher proportion of unsaturated FAs（C16：1 and C18：1）.However,when cells overexpressing DGAT2 were cultured with FA-DM for30 min,the FA composition was almost identical to that of controls.Further,the proportion of stable isotope-labeled FAs were similar in 3T3-L1 adipocytes overexpressing DGAT2 and control cells cultured in IFA-DM for 12 d.These results collectively indicate that the higher proportion of mono-unsaturated FAs,C16：1 and C18：1,may originate from de novo FA synthesis but not from the uptake of specific FAs from the medium.This hypothesis is further supported by evidence that both mRNA and protein expression of genes involved in FA synthesis（ACACA,FASN,SCD1,and A-FABP?     

Dietary essentiality of ascorbic acid in rohu larvae: Quantification with ascorbic acid enriched zooplankton

Carp larvae, like any other fish larvae dependon natural food during first few days of theirlife. In nursery conditions, high mortality andslow larval growth are of common occurrence;sub-optimal nutrition might be a possiblereason for such consequences. To improve thesituation the effect of feeding ascorbicacid-enriched live food on survival, growth,tissue biochemical composition includingascorbate level was evaluated in first feeding(3 days old) larvae (av. wt. 2.2 mg) of therohu carp, Labeo rohita (Ham.) for aperiod of 15 days (temp. 28.6 ± 1 °C)under natural photoperiod. The larvae (stockingdensity 10 l^–1) were offered enriched andnon-enriched zooplankton ad libitumfollowing a rigid schedule with four feedingregimes, each having 3 replicates. In treatmentT₁, non-enriched zooplankton (Moina,Daphnia, Cyclops, Diaptomus) and in T₂,T₃, T₄ ascorbic acid enriched (12 henrichment) zooplankton [@10%, 20% and 30%ascorbyl palmitate (AP) inclusion in diet ofzooplankton] were offered. Highest survival(90%) and growth (9563% live weight gain)could be seen in T₃ group and the lowestin T₁ (62% survival and 805% live weightgain), thus confirming the dietary essentialityof ascorbic acid for rohu larvae. Therequirement has been shown to be 1409 µg/gdry diet. Whole body tissue analyses for crudeprotein, total lipid and RNA: DNA ratiofollowed the same trend as that of growthresponse and percent survival. Significantpositive correlation (r = 0.949 and 0.861) couldbe found with muscle RNA/DNA ratio and muscleRNA content with specific growth rate indifferent treatments. Significant differencewas found in tissue ascorbate levels betweenenriched plankton fed groups, being highest in T₃. Such live foodmediated vitamin transfer might be an effectivemeans to provide higher plane of nutrition forhigh survival and rapid growth for rohu larva.     

Artemia prey size and mode of presentation: Effects on the survival and growth of phyllosoma larvae of southern rock lobster (Jasus edwardsii)

Performance of phyllosoma of thesouthern rock lobster (Jasus edwardsii)was examined after feeding Artemia-baseddiets. Survival and growth of newly-hatchedlarvae cultured to Stage III were lower(p < 0.05) when fed 0.8 mm Artemia than1.5 mm or 2.5 mm Artemia alone or 1.5 mmArtemia in combination with pieces ofmussel (Mytilus edulis planulatus) gonad.This could not be attributed to deficiencies inthe composition of fatty acids but appeared tobe due to the inability of larvae to capturesufficient appropriate-sized, enrichedArtemia for their nutritional requirements.There was an indication that survival andgrowth were higher between Stages III and Vwhen fed 2.5 mm Artemia than 1.5 mmArtemia alone or in combination with musselpieces. However, Stage VI larvae grew to asimilar size at Stage VIII when fed 1.5 mm or2.5 mm Artemia. Unexpectedly, larvae fedthe combination of 1.5 mm Artemia plusmussel supplement had lower survival than foundpreviously, and generally lower than when fed 1.5 mm Artemia alone. This was despitean apparent nutritional profile (lipid contentand fatty acid composition) of mussel more akinto that of newly-hatched phyllosoma thanenriched Artemia. On the other hand,survival and growth to Stage VIII were higherwhen larvae were fed alginate pelletscontaining Artemia than when fed 1.5 mmor 2.5 mm Artemia alone.     

Dietary inclusion of crystalline D- and L-methionine: effects on growth, feed and protein utilization, and digestibility in small and large Atlantic salmon (Salmon salar L.)

Use of crystalline methionine to optimize amino acid profiles in feeds for Atlantic salmon (Salmo salar L.) was studied in three experiments. A basic experimental diet with 25% of dietary protein from marine origin and the remaining 75% from soya protein concentrate was used either with or without supplementation of L ‐methionine, D ‐methionine or DL ‐methionine. A fishmeal‐based diet was used as control. In experiment 1, growth performance of 100 g salmon in seawater was improved by supplementation with the different methionine products. Growth performance was, however, inferior to that of fish fed the control. Feed conversion ratios (FCR) exhibited similar trends to specific growth rate (SGR) with methionine supplementation, but again not as the control. Protein retention, measured as productive protein value (PPV), was significantly better in the control and D ‐methionine groups as compared with either L ‐methionine or the nonsupplemented group. These trends were confirmed in experiment 2, using fish of the same size, where an absorption and digestibility study of D ‐ and L ‐methionine clearly indicated that D ‐methionine was absorbed as well as L ‐methionine. Results indicated that racemization of D ‐ into L ‐methionine occurs in liver and kidney in Atlantic salmon. Ornithine decarboxylase (ODC) activity in liver was significantly affected by dietary treatment and there was a good correlation with growth and feed utilization data. In experiment 3, a growth study with large salmon (1 kg) was performed using increasing supplementation of DL ‐methionine to the basic diet. Results indicated a methionine requirement for maximum growth of 9.2 g kg^–1 feed dry matter (DM) kg^–1 growth. This is slightly higher than that reported in the literature. The present study clearly shows that optimizing the amino acid profile by inclusion of a single crystalline amino acid (crystalline methionine) does not give the same protein utilization as that of a diet in which the amino acids are mainly protein bound. D ‐methionine was found to result in better protein utilization than L ‐methionine when measured as PPV.     

Effects of dietary thiamin on the physiological status of the grouper <Emphasis Type="Italic">Epinephelus coioides</Emphasis>

This study was conducted to evaluate the effects of dietary thiamin on the physiological status of the juvenile grouper, Epinephelus coioides. Graded levels of thiamin (0.08, 0.50, 2.12, 3.15, 4.63, 12.37 mg thiamin kg⁻¹ diet) were fed to grouper juveniles (mean weight: 16.97 ± 0.14 g) for 10 weeks. Although fish fed the thiamin-deficient (TD) diet showed no obvious symptoms of thiamin deficiency or increased mortality, those fed the lowest doses of thiamin (0.08 and 0.50 mg thiamin kg⁻¹ diet) had significantly decreased transketolase activity in the liver. In addition, the level of liver thiobarbituric acid reactive substances in fish fed the TD diet was 33–67% higher than that in fish with the thiamin-supplemented diet. There were no significant differences in superoxide dismutase activity between the different groups of fish.     

Effects of dietary lipid sources on growth and fatty acid composition of juvenile shrimp, Litopenaeus vannamei

This experiment was conducted to evaluate the effects of dietary lipid sources on the growth performance and fatty acid (FA) composition of juvenile shrimp, Litopenaeus vannamei. Six isoenergetic and isonitrogenous semi‐purified diets containing casein, solvent‐extracted soybean meal and gelatin as protein sources, were supplemented with 60 g kg⁻¹ of lipid sources. The lipid sources included: pollack fish oil (PO), pork lard (PL), soy oil (SO), peanut oil (PN), rapeseed oil (RO) and a mixture of pollack fish oil and soy oil (POSO 1 : 1 w/w). Each diet was fed to juvenile shrimp (0.10 g average weight) four times daily in triplicate tanks to apparent satiation (feeding ratio was about 8%) for 8 weeks. At the end of the experiment, weight gain, specific growth rate and protein efficiency ratio were significantly higher for shrimp fed the diet containing PO and the POSO mixture oil than the other lipid sources. The nutritional values of SO, RO, PN and PL were similar. Shrimp fed on PO, mixture oil of POSO and SO had better survival rates than the other lipid sources, and shrimp fed the PL had the lowest survival rate. There were significant differences in lipid contents of whole body and hepatopancreas amongst the dietary treatments; however, lipid contents of tail muscle were not significantly affected by the dietary lipid sources. Shrimps fed POSO diet had higher protein content in whole body than those fed the other lipid sources, and shrimp fed PO diet had highest crude protein content of the tail muscle. A high correlation was found between dietary FA composition and FA composition of whole shrimp. FA composition of the whole body was generally affected by dietary lipid sources, especially dietary unsaturated FA.     

Amino acid composition of Arctic charr, Salvelinus alpinus (L.) and the prediction of dietary requirements for essential amino acids

Embryo somatic tissues, non‐somatic yolk‐sac materials, and whole, individual fingerlings (age 0+) of Arctic charr, Salvelinus alpinus (L.), as well as a commercial trout diet, were analysed for a wide spectrum of amino acids. Analytical material consisted of prefeeding swim‐up fry that were separated into discrete yolk sac and somatic embryo tissue samples. Amino acid concentrations in fry somatic tissue and whole fingerlings were generally very similar to each other, but were lower than those measured in yolk materials. Higher correlations were observed between the majority of specific amino acid concentrations in the trout diet when compared with fingerling data (r² = 0.91) and fry somatic tissue data (r² = 0.89), than when correlated with fry yolk sac material (r² = 0.76). These results indicate that the essential amino acid profiles of fry somatic tissue and whole fingerlings are closer to that of a commercial feed than they are to the endogenous profiles found in the embryonic yolk sac material itself. The dietary ratios of individual essential amino acids were also compared with the total essential amino acid concentrations (A/E ratios) in whole fingerling tissues, and these ratios could be used to accurately estimate the apparent essential amino acid requirements of Arctic charr. The rationale for using carcass amino acid composition data to estimate the dietary essential amino acid requirements of Arctic charr is discussed.