Comparison of muscle fatty acid composition and lipid stability in lambs stall-fed or pasture-fed alfalfa with or without sainfoin pellet supplementation.

Currently, consumers are increasingly interested in obtaining high-quality and healthy lamb meat. Compared to grain-based diets, dietary forage legumes such as alfalfa and condensed tannin (CT)-rich sainfoin increase the levels of polyunsaturated fatty acids (PUFAs) that are beneficial for health in lamb meat thanks to their high content in PUFA and/or their impact on ruminal biohydrogenation. However, they can therefore adversely affect its oxidative stability. Thus, the impact of dietary forage legumes on lamb longissimus thoracis (LT) muscle FA composition and their stability to peroxidation was studied in 36 Romane lambs grazing alfalfa (AF; n = 12) or alfalfa plus daily supplementation with CT-rich sainfoin pellets (AS; n = 12; 15 g DM/kg BW, 42 g CT/kg DM) or stall-fed concentrate and grass hay indoors (SI; n = 12). Lambs were slaughtered at a mean age of 162 ± 8.0 days after an average experimental period of 101 ± 8.1 days. Forage legumes-grazing lambs outperformed SI lambs in LT nutritional quality, with more conjugated linoleic acids and n-3 PUFAs, especially 18:3n-3, eicosapentaenoic and docosahexaenoic acids (P < 0.001), and thus lower n-6 PUFA/n-3 PUFA and 18:2 n-6/18:3 n-3 ratios (P < 0.001). Peroxidizability index was higher (P < 0.001) in LT muscle of forage legumes-grazing lambs. Concurrently, two endogenous antioxidant enzyme activities, superoxide dismutase and glutathione peroxidase, were, respectively, similar and lower (P < 0.001) for forage legumes-grazing compared with SI lambs. A lower vitamin E level in SI lambs compared with forage legumes-grazing lambs (1.0 v. 3.8 mg/g, P < 0.001) could explain that malondialdehyde content, a marker of lipid oxidation intensity, was 0.63 µg/g in SI after 8 days in aerobic packaging conditions, whereas it remaining steady at 0.16 µg/g in forage legumes-grazing lambs. Dietary forage alfalfa thus improved FA composition of lamb LT muscle and their stability to oxidation when compared to SI lambs. However, supplementation of alfalfa-grazing lambs with CT-rich sainfoin pellets did not affect the nutritional quality of LT muscle FAs.

Are there consistent relationships between major connective tissue components, intramuscular fat content and muscle fibre types in cattle muscle?

Intramuscular connective tissue (IMCT) is mainly composed of several fibrils (known as total collagen (TCol)) linked between each other by different chemical cross-links (CLs), the whole being embedded in a matrix of proteoglycans (PGs). In the field of beef quality, there is limited information on the role of CLs and PGs. Accordingly, several authors suggest that, to investigate the role of IMCT, it is important to investigate them just like TCol and insoluble collagen (ICol). In muscle, there are two other components, the muscle fibres and intramuscular fat (IMF) content. There are limited data on the relationships between these three components of muscle and then on possibility to independently manipulate these characteristics in order to control the final quality of meat. The present study aimed to investigate whether consistent relationships exist between these different components of muscle. Therefore, the present study compared four muscles of two cattle types (dairy and beef) to determine associations between TCol, ICol, CLs and PGs. Data were analysed across and within muscle (M) and animal type (AT) based on residuals. There was a strong M and AT effect for all muscle characteristics and an interaction M × AT for type I muscle fibres and IMF. Correlations between TCol, ICol and their CLs were M- and AT-independent. Total proteoglycans were positively correlated with TCol and ICol in a muscle-dependent manner irrespective of AT, but no correlation was found with CLs. On the contrary, CLs were negatively correlated with the ratio TPGs : TCol in an M-dependent manner, irrespective of AT. TCol, ICol and CLs were positively and negatively correlated with type IIA and IIB+X muscle fibres only in longissimus thoracis (LT) muscle, regardless the AT. Insoluble collagen was the only parameter of IMCT to be correlated with type I muscle fibres but only in LT muscle, irrespective of AT. There was no correlation between PGs and muscle fibre types, but PGs were the only IMCT component to be related with IMF in an M-dependent manner, irrespective of AT. Finally, there was no correlation between muscle fibre types and IMF content within M and AT. This study revealed that there is a strong relationship between IMCT components irrespective of M, an M-dependent relationship between the IMCT components and muscle fibre types and few (only with PGs) or no relationship between IMF and IMCT and muscle fibres.

Prediction of beef meat fatty acid composition by visible-near-infrared spectroscopy was improved by preliminary freeze-drying.

The aim of this study was to compare visible-near-infrared spectroscopy (VIS/NIRS) models developed from fresh or freeze-dried samples for predicting the fatty acid (FA) composition of beef samples. The hypothesis tested is that the removal of water from samples could improve the VIS/NIRS model performance. A total of 454 beef samples obtained from different bovine muscles were used. No significant differences were found in the performance of VIS/NIRS models developed from fresh or freeze-dried samples for predicting both major individual FAs and families of FAs and for some FAs (16:0, 18:0, 18:1 n-9, 18:2 n-6, 20:4 n-6, 22:5 n-3, 22:6 n-3, saturated, mono-unsaturated FA, and total n-3 long chain poly-unsaturated FAs (PUFA)). In contrast, the standard error of predictions for total PUFAs, total n-3 PUFAs, total conjugated linoleic acid, 20:5 n-3, and 18:3 n-3 were improved (by 21% on average; P < .05) in freeze-dried samples compared with fresh samples.

Performance of fluorescence spectroscopy for beef meat authentication: Effect of excitation mode and discriminant algorithms.

This study evaluated the performance of classical front face (FFFS) and synchronous (SFS) fluorescence spectroscopy combined with Partial Least Square Discriminant Analysis (PLSDA), Support Vector Machine associated with PLS (PLS-SVM) and Principal Components Analysis (PCA-SVM) to discriminate three beef muscles (Longissimus thoracis, Rectus abdominis and Semitendinosus). For the FFFS, 5 excitation wavelengths were investigated, while 6 offsets were studied for SFS. Globally, the results showed a good discrimination between muscles with Recall and Precision between 47.82 and 94.34% and Error ranging from 6.03 to 32.39%. For the FFFS, the PLS-SVM with the 382nm excitation wavelength gave the best discrimination results (Recall, Precision and Error of 94.34%, 89.53% and 6.03% respectively). For SFS, when performing discrimination of the three muscles, the 120nm offset gave the highest Recall and Precision (from 57.66% to 94.99%) and the lowest Error values (from 6.78 to 8.66%) whatever the algorithm (PLSDA, PLS-SVM and PCA-SVM).

Factors influencing proportion and composition of CLA in beef.

Bovine meat is criticised for the bad nutritional image of its lipids and fatty acids. However, with dairy products, beef is the major source of conjugated linoleic acid (CLA) which could have several human health benefits. The present study compared, from data of five nutritional experiments on bovine animals performed by the laboratory, the impact of factors linked to the animals (breed, age, sex, type of muscle) and to feeding conditions (basal diet, lipid supplements) on the CLA proportion and composition in muscles. Among these factors, linseed supplementation was an efficient way to increase CLA proportion in beef (+22% to +36%) but was highly modulated by the nature of the basal diet, and by intrinsic factors (breed, age/sex, type of muscle) since these ones could modulate CLA proportion in beef from 24% to 47%. Moreover, these factors modified also the proportion of cis,trans-CLA, related to cis,cis- and trans,trans-isomers. Specific biological properties of these latter isomers should be determine to understand the consequences of intramuscular CLA isomer variations for the health of consumers.

Potential of fluorescence spectroscopy to predict fatty acid composition of beef.

The present study aimed to evaluate and compare the ability of front face (FFFS) and synchronous fluorescence spectroscopy (SFS) to predict total fat and FA composition of beef LT muscles coming from 36 animals of 3 breeds (Angus, Limousin and Blond d'Aquitaine). The regression models were performed by using Partial Least Square (PLS) method. In spite of the low number of samples used, the results of this preliminary study demonstrated the ability of fluorescence spectroscopy to predict meat lipids. Nonetheless, the results suggested that the fluorescence spectroscopy is more suited to measure SFA (R(2)p≥0.66; RPD≥2.29) and MUFA (R(2)p≥0.48; RPD≥1.49) than PUFA (R(2)p≤0.48; RPD≤1.63). Moreover, R(2) and RPD factors obtained with FFFS were greater compared to the ones obtained with SFS suggesting that FFFS is more adapted to measure lipid composition of beef meat.

An innovative approach combining Animal Performances, nutritional value and sensory quality of meat.

This work sets out a methodological approach to assess how to simultaneously control together Animal Performances, nutritional value, sensory quality of meat. Seventy-one young bulls were characterized by 97 variables. Variables of each element were arranged into either 5 homogeneous Intermediate Scores (IS) or 2 Global Indices (GI) via a clustering of variables and analysed together by Principal Component Analysis (PCA). These 3 pools of 5 IS (or 2 GI) were analysed together by PCA to established the links existing among the triptych. Classification on IS showed no opposition between Animal Performances and nutritional value of meat, as it seemed possible to identify animals with a high butcher value and intramuscular fat relatively rich in polyunsaturated fatty acids. Concerning GI, the classification indicated that Animal Performances were negatively correlated with sensory quality. This method appeared to be a useful contribution to the management of animal breeding for an optimal trade-off between the three elements of the triptych.

Breeds and muscle types modulate performance of near-infrared reflectance spectroscopy to predict the fatty acid composition of bovine meat.

This study aims to assess near-infrared reflectance spectroscopy feasibility for predicting beef fatty acid (FA) composition. Experimental scheme included four breeds (Angus, Blond d'Aquitaine, Charolais, Limousin) and three muscles, Longissimus thoracis (LT), Rectus abdominis (RA), Semitendinosus (ST). The results showed that 1) increasing FA content variability with several breeds increased calibration model reliability (R(2)CV>0.86) for the major individual and groups of FA unless polyunsaturated FAs, 2) Longissimus thoracis FAs were better predicted than RA FAs while no ST FAs were correctly predicted (R(2)CV<0.71). This difference could be explained by FA content, FA variability or specific muscle physico-chemical characteristics.

Thyroid hormone synthesis in thyroglobulin secreted by porcine thyroid cells cultured on porous bottom chambers. Effect of iodide.

The long-term iodination of thyroglobulin secreted into the apical medium of thyroid cells cultured as monolayers on porous bottom chambers reached 5.87 +/- 1.66 atoms of iodine/mol thyroglobulin after 11 days incubation in the presence of TSH (0.1 mU/ml) and iodide (0.5 microM) in the basal medium. This iodinated thyroglobulin contained thyroid hormones (T3 + T4) which involved 22.7% of the thyroglobulin iodine content. The iodoamino acid content was, in residues per mole, 2.2 +/- 0.35 for monoiodotyrosine, 0.74 +/- 0.04 for diiodotyrosine, 0.23 +/- 0.04 for T4, and 0.098 +/- 0.02 for T3. Kinetic studies showed that a minimal level of iodination (2.05 +/- 0.26 atoms iodine/mol thyroglobulin) was necessary for hormonogenesis. A maximal level of iodination and hormonogenesis was obtained with 0.5 microM iodide added daily to the basal medium. In these conditions, hormonogenesis efficiency reached about 40% (a value close to this one observed in vivo). Above 0.5 microM iodide, both iodination and T4 synthesis were inhibited (28.3% and 73.9%, respectively, for 1 microM iodide). Our culture system makes it possible to demonstrate that this high iodide concentration in the basal medium did not increase apical iodide concentration above 10 microM but decreased apical thyroglobulin concentration. The inhibitory effect of iodide on hormonogenesis cannot be due to a competition with tyrosine residues of thyroglobulin for their binding to thyroperoxidase although it could be related, at least in part, to a decrease in protein synthesis.

Glycosylation of thyroglobulin secreted by porcine cells cultured in chamber system: thyrotropin controls the number of oligosaccharides and their anionic residues.

Porcine thyroid cells were cultured in porous bottom chambers in the presence or in the absence of TSH added to the basal medium. Radiolabeled-sugar (3H-mannose) was added to the basal medium on day 11 for 4 days and the glycosylation of thyroglobulin (Tg), the major glycoprotein secreted into the apical medium, was studied. The incorporation of 3H-mannose per molecule of Tg was increased 1.5-fold by a 50 microU/ml minimal concentration of TSH. The distribution of glycopeptides (after pronase digestion) on concanavalin A sepharose column was not modified by the presence of TSH. However this distribution was different from that observed for Tg extracted from gland (more multiantennary units than biantennary units and polymannose units). After desialylation and desulfation, the sizes of the oligosaccharide chains analyzed on HPLC appeared similar when cells were cultured under stimulation or not. Thus TSH enhanced sugar incorporation without modifying either the distribution of the different oligosaccharide moieties or their sizes. Consequently the effect of TSH was a 1.5-fold increase in oligosaccharide chains linked to asparagine residues. 3H-Mannose-oligosaccharide chains were then analyzed on ion-exchange HPLC before and after desialylation and desulfation. The number of anionic residues per oligosaccharide unit (particularly sulfate residues) was higher in the absence of TSH than in the presence of TSH. Nevertheless, since TSH increased the number of carbohydrate units per molecule of Tg 1.5-fold, the total content of anionic residues bound to oligosaccharide units per molecule of Tg seems not to be modified by TSH.

Thyrotrophin regulation of apical and basal exocytosis of thyroglobulin by porcine thyroid monolayers.

Exocytosis, the ultimate step in thyroglobulin secretion, has been studied in porcine thyroid cells cultured in monolayers on the permeable bottom of culture chambers. We have previously demonstrated, using this culture system, that apical secretion accounts for 85-95% of total secretion of newly synthesized thyroglobulin. When cells were cultured for several days with bovine TSH (25 microU/ml) in the basal medium, the rate of glycoprotein accumulation in the upper compartment was three times higher than that in the absence of TSH. In contrast, the rate of thyroglobulin released into the basal medium (5-15% of total secreted thyroglobulin) appeared unmodified by chronic TSH stimulation. To investigate the effect of acute TSH stimulation on thyroglobulin exocytosis in the apical and basal compartments, pulse-chase experiments were carried out with the same culture system. The release of radiolabelled thyroglobulin (1.5-h pulse) into the apical medium was increased threefold during the 2-h chase period under TSH stimulation. The radiolabelled thyroglobulin released into the basal medium was increased only 1.5- to 2-fold, and stimulation disappeared after 1 h. The effect of TSH was maximal when the chase medium contained 50 microU TSH/ml. However, cells cultured for several days in the presence of 25 microU TSH/ml before the pulse-chase experiment, appeared desensitized to acute TSH stimulation. Similar responses were observed when the chase medium contained 8-chloro-cyclic AMP or cholera toxin. This study provides another example of the pleiotropic effect of TSH, mediated by cyclic AMP, on the sequential steps of thyroglobulin gene expression in cultured thyroid cells in which the polar character of the epithelial cells is well preserved.

Long-term iodination of thyroglobulin by porcine thyroid cells cultured in porous-bottomed culture chambers: regulation by thyrotrophin.

Thyroid cells cultured as monolayers on the porous bottom of culture chambers have been shown to express some specific functions of thyroid follicles. This system, which allows independent access to apical and basal media, is suitable for the long-term study of polarized processes, as the cells maintain their polarized organization. Iodination of thyroglobulin has been investigated under different culture conditions in the presence or absence of TSH. Apical thyroglobulin accumulation, apical iodide concentration and thyroglobulin iodination have been followed simultaneously. Iodide (0.5 mumol/l) was added to basal medium at various stages: only once for 4-day incubations and at each medium change or daily for longer experiments. TSH increased the amount of thyroglobulin secreted into the apical medium by five- to sixfold, whereas high basal iodide concentrations (greater than 5 mumol/l) inhibited thyroglobulin secretion by TSH-stimulated cells. TSH increased iodide uptake giving an iodide concentration ratio between apical and basal media of about 5. Thyroglobulin iodination was dependent upon TSH. Thyroglobulin was iodinated only in the apical compartment. Secretion and iodination of thyroglobulin were polarized phenomena, but the polarity of iodination was total whereas the polarity of secretion was only partial (10% basal secretion). This functional asymmetry was maintained for up to 29 days. The maximal incorporation of iodine into thyroglobulin obtained was never higher than 3.5 atoms/mol. Apical iodide concentrations from 1 to 15 mumol/l, depending on culture conditions, did not increase this value. These results suggest that cells cultured in this culture system are able to reproduce several steps of thyroidal iodide metabolism although there may be unknown factors which could interfere and reduce the efficiency of thyroglobulin iodination.

Phenol red: an inhibitor of thyroglobulin iodination in cultured porcine thyroid cells.

Phenol red, commonly used as a pH indicator in tissue culture media, is known to possess estrogenic properties. We investigated the effect of phenol red on the process of thyroglobulin iodination which occurs only at the apical surface of porcine thyroid cells when cultured in porous bottom chambers. When phenol red was added simultaneously to both compartments (apical and basolateral), separated by the polarized monolayer, thyroglobulin iodination was inhibited by about 86% without any effect on thyroglobulin secretion and apical iodine concentration. When phenol red was added separately to either the apical or basal media, inhibition was 68% and 43%, respectively. A large amount of phenol red which was introduced into the basal medium crossed through the monolayer. Thus, inhibition was dependent upon the concentration of phenol red present in the apical compartment. A maximal inhibition was observed from 30 microM apical concentration. Phenol red acts as a substrate for thyroperoxidase in the iodination reaction.

Fatty acid metabolism and very low density lipoprotein secretion in liver slices from rats and preruminant calves.

The liver of bovine animals possesses a low ability to secrete triglycerides (TG) as part of the very low density lipoproteins (VLDL) compared with rat liver. We compared hepatic fatty acid (FA) metabolism between rat and calf in order to determine the limiting steps of TG-VLDL secretion in bovine animals. Liver slices from young Sprague-Dawley rats and preruminant Holstein x Friesian calves were incubated for 7 h with increasing concentrations (0.1, 0.2, 0.4, and 0.8 mM) of [14C]oleate. The oxidation of oleate to CO2 and acid-soluble products was 2- to 3-fold higher in rat than in calf liver slices. Since oleate uptake was 2-fold higher in rat than in calf, the oxidation rate represented 20-29% of oleate uptake in both animal species. Oleate was essentially incorporated into the neutral lipids (75-87% of total lipids) that were stored mainly in the cytosol in both animal species (81-90% of neutral lipids). The accumulation of neutral lipids in the cytosol was 3.4-fold higher while VLDL secretion was 6- to 18-fold more efficient in rat than in calf liver slices. Our results indicate that the slow rate of VLDL secretion by bovine liver is probably due to the limited availability of TG for VLDL packaging rather than to the preferential oxidation of FA.

Effects of fish oil and additional starch on tissue fatty acid profile and lipogenic gene mRNA abundance in lactating goats fed a diet containing sunflower-seed oil.

In dairy cattle, diet supplementation with oils affects the lipid metabolism in body tissues via changes in the partitioning and deposition of fatty acids (FAs) and lipogenic gene expression; however, limited data are available in goats. Eight Alpine goats were fed a grassland hay diet supplemented with 90 g/day of sunflower-seed oil or 90 g/day of sunflower-seed oil and fish oil (2 : 1) plus additional starch. The goats were slaughtered on day 21 of the treatments and samples of the mammary secretory tissue, liver, omental and perirenal adipose tissues (ATs) were collected to characterise their FA composition and the mRNA abundance of lipogenic genes and transcription factors involved in their regulation, and to examine the impact of the diet composition on the same parameters. The results are in agreement with the specific physiological adaptation in the lipid metabolism of body tissues that is likely to occur during late lactation because of the coexistence of an active lipogenesis in the mammary secretory tissue and a significant anabolic activity in the ATs. These latter tissues were characterised by high concentrations of saturated FA and very low polyunsaturated FA (PUFA) levels. The content of PUFA was relatively higher in the mammary secretory tissue, in particular in the case of polyunsaturated C18. The highest PUFA contents were found in the liver, in accordance with the greater mRNA abundances of the genes that encode the necessary enzymes for very long-chain n-3 and n-6 PUFA synthesis. However, substantial differences between n-3 and n-6 pathways would most likely exist in the goat liver. Overall, differences in diet composition induced limited changes in the mRNA abundance of genes involved in lipid metabolism, and these were not associated with the few variations observed in tissue FA composition.

A grass-based diet favours muscle n-3 long-chain PUFA deposition without modifying gene expression of proteins involved in their synthesis or uptake in Charolais steers.

N-3 long-chain polyunsaturated fatty acids (n-3 LCPUFA) are subject of growing interest as they are of particular relevance for meat quality and human health. However, their content in the muscles of cattle is generally low probably as the complex result of their biosynthesis from dietary n-3 PUFA in the muscle and/or in other tissues/organs and of their subsequent uptake by the muscle. In view of this, this study aimed at understanding whether the changes in the muscle n-3 LCPUFA content, depending on the diet (maize silage v. grass) or the muscle type (Rectus abdominis, RA v. Semitendinosus, ST) in 12 Charolais steers, were related to variations in the gene expression of proteins involved in n-3 LCPUFA biosynthesis or cellular uptake. Tissue fatty acid composition was analysed by gas-liquid chromatography and mRNA abundance of proteins by quantitative real-time PCR. The grass-based diet resulted in a 2.3-fold (P < 0.0002) increase in both RA and ST n-3 LCPUFA content compared with the maize silage-based diet, whereas no difference in the expression of genes involved in n-3 LCPUFA biosynthesis and uptake was observed between diets. ST exhibited a 1.5-fold higher n-3 LCPUFA content than RA (P < 0.003), whereas the gene expression of proteins involved in n-3 LCPUFA biosynthesis and uptake was 1.3- to 18-fold higher in RA than in ST (P < 0.05). In conclusion, diet- or muscle type-dependent changes in the muscle n-3 LCPUFA content of Charolais steers did not seem to be mediated by the gene expression regulation of proteins involved in the biosynthesis or uptake of these fatty acids.

Breed and dietary linseed affect gene expression of enzymes and transcription factors involved in n-3 long chain polyunsaturated fatty acids synthesis in longissimus thoracis muscle of bulls.

N-3 long-chain (LC) PUFA are known to be beneficial for human development and health. These properties explain the increasing interest in promoting n-3 LC PUFA deposition in bovine muscles, leading to healthier meats. In this context, this study aimed to identify possible limiting steps in the bioconversion of 18:3n-3 into n-3 LC PUFA in the longissimus thoracis (LT) muscle of 36 Aberdeen Angus, Limousin, and Blond d'Aquitaine bulls (n = 12 per breed) that were fed, for the 105-d finishing period, either a concentrate-based diet (25% molasses straw to 75% concentrate, on a raw basis; CON) or the same CON diet supplemented with extruded linseed (44.5 g lipid/kg diet DM) mixed into the concentrate (LINS). The fatty acid (FA) composition of the LT muscle was determined by GLC, and the mRNA abundances for enzymes and transcription factors involved in n-3 LC PUFA synthesis were determined by quantitative real-time PCR. The total lipid concentration in the LT muscle was approximately 2.4-fold greater (P < 0.001) in Angus bulls than in the other breeds and composed of the greatest n-3 PUFA content (P < 0.001) including 18:3n-3 (P < 0.001) and n-3 LC PUFA (P < 0.02), primarily 20:5n-3 (P < 0.007) and 22:5n-3 (P < 0.04). These data were associated with a lesser gene expression (P < 0.02) of 2 enzymes [acyl-CoA oxidase 1 (ACOX1) and L-bifunctional protein (L-PBE)] and 2 transcription factors [liver X receptors (LXR) α and ß] in the LT muscle of Angus bulls compared with gene expression in Limousin bulls. Moreover, the mRNA of elongase 5 was only present in trace amounts in the LT muscle of the 3 breeds. The addition of linseed to the diet resulted in greater deposition of 18:3n-3 (P < 0.001) in the LT muscles of the 3 breeds, without any major changes (P > 0.34) in the n-3 LC PUFA content. Dietary linseed stimulated (P < 0.04) the gene expression of all enzymes and transcription factors involved in n-3 LC PUFA synthesis except elongases 2 and 5 (P > 0.19), the expression of which remained weak and was not inducible. These results reveal a limited capacity for n-3 LC PUFA synthesis from 18:4n-3 (substrate of elongase 5) in the LT muscles of Blond d'Aquitaine, Limousin, and Angus bulls. Therefore, further investigations on the cellular regulation of elongase gene expression are needed to identify the physiological or nutritional factors that efficiently stimulate elongase expression in beef cattle.

Distinct metabolism of linoleic and linolenic acids in liver and adipose tissues of finishing Normande cull cows.

Feeding strategies based on the addition of plant lipids rich in polyunsaturated fatty acids (PUFAs) in diets of bovines during the finishing period are common to enhance the nutritional value of meat. However, following rumen biohydrogenations, these FAs could still be metabolised in various tissues/organs involved in the FA metabolism such as the liver and adipose tissues (ATs), thus affecting their subsequent deposition in muscles. In this context, the objective of this study was to characterise the various metabolic pathways of linoleic acid (LA) and α-linolenic acid (ALA) in the liver and ATs (subcutaneous (SC) and inter-muscular (IM)) of Normande cull cows fed a diet supplemented (LR) or not (C) with extruded linseeds and rapeseeds, using the ex vivo incubated tissue slice method. Hepatic uptake of both FAs was higher with the LR than with the C diet (P = 0.02). For the two diets, ALA uptake was higher than that of LA (+46%, P = 0.04). ALA was much more degraded by ß-oxidation (>50% of ALA present in cells) than LA (∼27%) with both diets (P = 0.015). Whatever the diet, ALA was not converted into longer and/or more unsaturated FA, whereas about 14% of LA was converted into 20:4n-6. The intensity of the esterification pathway was higher (+70%, P = 0.004) with the LR than with the C diet, for both FAs. Hepatic secretion of ALA as part of the very-low-density lipoprotein particles was lower than that of LA (-58% and -23% for C and LR diets respectively, P = 0.02). In SC and IM ATs, dietary lipid supplementation did not alter metabolic pathways of LA and ALA. They were efficiently taken up by ATs (>68% of FA present in the medium), with uptake being higher for IM than for SC AT (+12%, P = 0.01). Moreover, LA uptake by ATs was higher than ALA uptake (+10.7%, P = 0.027). Both FAs were mainly esterified (>97% of FA present in adipocytes) into neutral lipids (>85% of esterified FA). Around 9.5% of LA was converted into 20:4n-6, whereas only around 1.3% of ALA was converted into 20:5n-3. We concluded that, in our experimental conditions, liver was highly active in ALA catabolism limiting its subsequent deposition in muscles. However, bovine liver and ATs were inefficient at converting ALA into long-chain n-3 PUFA, but actively converted LA into 20:4n-6.

Nitrogen- and energy-imbalanced diets affect hepatic protein synthesis and gluconeogenesis differently in growing lambs.

The aim of this study was to assess the metabolic fate of AA (endogenous or export protein synthesis, gluconeogenesis, or oxidation) after an imbalanced supply of energy and N in the diet of growing lambs. Eighteen INRA 401 lambs (3 mo old, 29.7 +/- 0.45 kg of BW) were fed 3 experimental diets, one providing a N and energy supply according to recommended allowances (control), one with 23% less N supply relative to energy (LN), and one with 19% less ME supply relative to N (LE). Animals were assigned to 6 blocks of 3, with each animal receiving 1 of the 3 diets, and the animals from each block were slaughtered on the same day. Liver slices from these lambs were incubated in a minimum salt medium (Krebs-Henseleit) containing physiological concentrations of propionate and AA as energy and N sources, similarly across all 3 treatments. Protein synthesis (endogenous and export) using [U-(14)C]valine and [(35)S]methionine, gluconeogenesis from [1-(14)C]propionate and [U-(14)C]alanine, and oxidation were measured. A relative sparing of AA at the liver level was observed with the LN diet because of reduced urinary N (-42%, LN vs. control, P < 0.001). The AA were also directed toward anabolic purposes in the LN diet via an increased endogenous and total export protein synthesis (+51%, LN vs. control, P = 0.01; also observed for fibrinogen synthesis, but not for albumin or transferrin) associated with a tendency for increased gluconeogenesis from alanine (+58%, LN vs. LE, P = 0.08). On the other hand, the LE diet resulted in a marked loss of N in urine (+24%, LE vs. control, P < 0.05), but no notable effect of the LE diet was demonstrated for protein synthesis or gluconeogenesis ex vivo. These data demonstrate a more efficient utilization of AA for anabolic purposes in the lambs fed LN, probably via an activation of some AA transport systems, to address the shortage of nitrogenous nutrients in the LN diet. By contrast, no such adaptation occurred in the LE lambs, probably because the regulatory mechanisms that prevailed in this case were the nutrient supply or hormones, which were not altered in our ex vivo experimental model.

9 cis,11 trans conjugated linoleic acid (CLA) is synthesised and desaturated into conjugated 18:3 in bovine adipose tissues.

Although endogenous synthesis of conjugated linoleic acid (CLA) in the mammary gland of lactating cows has been already well documented, no study has determined so far as to which tissue and/or organ is involved in CLA synthesis in the growing ruminant except one study showing that CLA synthesis does not occur in ruminant liver. In this context, adipose tissue appears to be a good candidate for endogenous synthesis of CLA in the growing ruminant. The aim of this study was to compare the respective metabolisms of 11trans 18:1 (vaccenic acid, VA) and 9cis,11trans 18:2 (rumenic acid) to that of stearic acid (the preferential substrate of Δ9 desaturase) in adipose tissues (subcutaneous, SC and intermuscular, IM) of six Charolais steers by using the in vitromethod of incubated tissue slices. Samples of SC and IM adipose tissues were incubated at 37°C for 16 h under an atmosphere of 95% O2/5% CO2 in a medium supplemented with 0.75 mM of fatty acid (FA) mixture (representative of circulating non-esterified FA) and 186 µM [1-14C]-18:0 or 58.6 µM [1-14C]-VA or 56 µM [1-14C]-9cis,11trans CLA. Viability of explants was verified by measuring metabolic functions (glucose uptake and glucose-6-phosphate dehydrogenase activity). After 16 h of incubation, FA uptake was similar for all FA (18:0, VA and 9cis,11trans 18:2) in both SC and IM adipose tissues (around 40%). Once in adipose tissue, all FA were preferentially esterified (>80% of cell FA) favouring neutral lipid synthesis (around 90% of esterified FA). Stearic acid was highly (27%) desaturated into oleic acid in SC adipose tissue whereas this desaturation was much lower (6.8%) in IM adipose tissue (P < 0.0001). VA was desaturated into 9cis,11trans CLA at a low extent of about 2.5% to 4.4% in both adipose tissues probably because of a limited affinity of Δ9 desaturase for VA. 9cis,11trans CLA was itself converted by desaturation into 6cis, 9cis,11trans 18:3 at the intensity of 10.8% and 14.5% of cell 9cis,11trans CLA in SC and IM adipose tissues, respectively. In conclusion, bovine adipose tissues of the growing ruminant were especially involved in the endogenous synthesis of CLA from VA and in its desaturation into conjugated derivative, mainly 6cis, 9cis,11trans 18:3, of which biological properties need to be elucidated.