Liver metabolism traits in two rabbit lines divergently selected for intramuscular fat

Intramuscular fat (IMF) has a large effect in the sensory properties of meat because it affects tenderness, juiciness and flavour. A divergent selection experiment for IMF in longissimus dorsi (LD) muscle was performed in rabbits. Since liver is the major site of lipogenesis in rabbits, the objective of this work is to study the liver metabolism in the lines of the divergent selection experiment. Intramuscular fat content, perirenal fat weight, liver weight, liver lipogenic activities and plasma metabolites related to liver metabolism were measured in the eighth generation of selection. Direct response on IMF was 0.34 g/100 g of LD, which represented 2.7 SD of the trait, and selection showed a positive correlated response in the perirenal fat weight. High-IMF line showed greater liver size and greater liver lipogenic activities of enzymes glucose-6-phosphate dehydrogenase and malic enzyme. We did not find differences between lines for fatty acid synthase lipogenic activity. With regard to plasma metabolites, low-IMF line showed greater plasma concentration of triglycerides, cholesterol, bilirubin and alkaline phosphatase than high-IMF line, whereas high-IMF line showed greater albumin and alanine transaminase concentrations than low-IMF line. We did not observe differences between lines for glucose, total protein and plasma concentrations. Phenotypic correlations between fat (IMF and perirenal fat weight) and liver traits showed that liver lipogenesis affects fat deposition in both, muscle and carcass. However, the mechanisms whereby liver lipogenesis affected IMF content remain to be clarified.     

Effect of sex,dietary glycerol or dietary fat during late fattening,on fatty acid composition and positional distribution of fatty acids within the triglyceride in pigs

The effect of sex, source of saturated fat (lard v. palm oil) and glycerol inclusion in the fattening diet on composition and fatty acid positional distribution in the triglyceride molecule was studied in pigs from 78 to 110 kg BW. Average daily gain and carcass characteristics, including ham and loin weight, were not affected by dietary treatment but sex affected backfat depth (P<0.01). A significant interaction between sex and glycerol inclusion was observed; dietary glycerol increased lean content in gilts but not in barrows (P<0.05 for the interaction). Individual and total saturated fatty acid (SFA) concentrations were greater in barrows than in gilts. In contrast, the concentration of total polyunsaturated fatty acids (PUFA) and of C18:2n-6, C18:3n-3, C20:3n-9 and C20:4n-6 in the intramuscular fat (IMF) was higher (P<0.05) in gilts than in barrows. Sex did not affect total monounsaturated fatty acids (MUFA) concentration in the IMF. The proportion of SFA in the subcutaneous fat (SF) was higher in barrows than in gilts (P<0.001). Within the individual SFA, sex affected only the concentrations of C14:0 and C16:0 (P<0.001). Dietary fat did not affect total SFA or PUFA concentrations of the IMF but the subcutaneous total MUFA concentration tended to be higher (P=0.079) in pigs fed lard than in pigs fed palm oil. Dietary glycerol increased total MUFA and C18:1n-9 concentration in the IMF and increased total MUFA and decreased C18:2n-6, C18:3n-3 and total PUFA concentrations in the SF. The data indicate that altering the fatty acid composition of the triglyceride molecule at the 2-position, by dietary intervention during the fattening phase, is very limited.     

Genomic regions influencing intramuscular fat in divergently selected rabbit lines

Intramuscular fat (IMF) is one of the main meat quality traits for breeding programmes in livestock species. The main objective of this study was to identify genomic regions associated with IMF content comparing two rabbit populations divergently selected for this trait, and to generate a list of putative candidate genes. Animals were genotyped using the Affymetrix Axiom OrcunSNP Array (200k). After quality control, the data involved 477 animals and 93 540 SNPs. Two methods were used in this research: single marker regressions with the data adjusted by genomic relatedness, and a Bayesian multiple marker regression. Associated genomic regions were located on the rabbit chromosomes (OCU) OCU1, OCU8 and OCU13. The highest value for the percentage of the genomic variance explained by a genomic region was found in two consecutive genomic windows on OCU8 (7.34%). Genes in the associated regions of OCU1 and OCU8 presented biological functions related to the control of adipose cell function, lipid binding, transportation and localisation (APOLD1, PLBD1, PDE6H, GPRC5D and GPRC5A) and lipid metabolic processes (MTMR2). The EWSR1 gene, underlying the OCU13 region, is linked to the development of brown adipocytes. The findings suggest that there is a large component of polygenic effect behind the differences in IMF content in these two lines, as the variance explained by most of the windows was low. The genomic regions of OCU1, OCU8 and OCU13 revealed novel candidate genes. Further studies would be needed to validate the associations and explore their possible application in selection programmes.     

The use of stoned olive cake and rolled linseed in the diet of intensively reared lambs: effect on the intramuscular fatty-acid composition

The aim of the present study was to evaluate the effect of the inclusion of stoned olive cake and rolled linseed in a concentrate-based diet for lambs on the fatty-acid composition of polar and non-polar intramuscular lipids of the longissimus dorsi muscle. To achieve this objective, 32 Appenninica lambs were randomly distributed into four groups of eight lambs each and were fed conventional cereal-based concentrates (diet C); concentrates containing 20% on a dry matter (DM) basis of rolled linseed (diet L); concentrates containing 35% DM of stoned olive cake (diet OC); and concentrates containing both rolled linseed (10% DM) and stoned olive cake (17% DM; diet OCL). The concentrates were administered together with grass hay at a 20:80 forage:concentrate ratio. Growing performances and carcass traits were evaluated. The fatty-acid composition was analysed in the total intramuscular lipids, as well as in the polar and neutral lipids. The average feed intake and the growth performance of lambs were not affected by the dietary treatments, as a consequence of similar nutritional characteristics of the diets. The inclusion of rolled linseed in the L and OCL diets increased the content of C18:3 n-3 in intramuscular total lipids, which was threefold higher in meat from the L lambs and more than twofold higher in meat from the OCL lambs compared with the C and OC treatments. The n-6:n-3 ratio significantly decreased in the meat from lambs in the L and OCL groups, reaching values below 3. The L treatment resulted in the highest level of trans-18:1 fatty acids in the muscle. Regardless of the dietary treatment, the t10-18:1 was the major isomer, representing 55%, 45%, 49% and 45% of total trans-18:1 for C, L, OC and OCL treatments, respectively. Neutral lipids from the OC-fed lambs contained the highest amount of c9-18:1 (more than 36% of total fatty acids); however, the content of c9-18:1 did not differ between the OC and C lambs, suggesting an intensive biohydrogenation of dietary c9-18:1 in the case of OC treatment. The highest content of c9,t11-18:2 was detected in the intramuscular fat from the L-fed lambs, followed by the OCL treatment. A similar trend was observed in the neutral lipid fraction and, to a lower extent, in the polar lipids.     

Effect of botanical composition of silages on rumen fatty acid metabolism and fatty acid composition in longissimus muscle and subcutaneous fat of lambs

To study the effect of feeding silages with different botanical composition, on rumen and lamb fat, 30 male lambs were assigned to five different silage groups for 11 weeks: botanically diverse silage (BDS); white clover silage (WCS); red clover silage (RCS), intensive English ryegrass silage (IRS) and crushed linseed and maize silage (MSL). Besides the silages, animals received organic wheat and barley and the MSL group additionally received bicarbonate (15 g/day). Silages were sampled when the bales were opened and analysed for fatty acid (FA) content and chemical composition. At slaughter, ruminal contents were sampled and 24 h after slaughter, longissimus muscle and subcutaneous (SC) fat were sampled. All samples were analysed for FA composition. The MSL group ingested the highest amount of FA (35.8 g/day v. 13.5, 19.4, 17.2 and 30.4 g/day for MSL v. BDS, WCS, RCS and IRS, respectively) and the sum of the major polyunsaturated FA, C18:2 n-6 and C18:3 n-3, was similar for groups BDS, WCS, RCS and MSL (61.3 g/100 g, 62.3 g/100 g, 62.3 g/100 g, 63.7 g/100 g of FA methylesters (FAME), respectively), while group IRS ingested higher proportions of these FA (74.5 g/100 g of FAME). Rumen data showed that animals fed BDS presented higher proportions of biohydrogenation intermediates, particularly C18:1 t11 and CLA c9t11, suggesting partial inhibition of rumen biohydrogenation. In the MSL group, the content of C18:3 n-3 in the rumen was highest, most probably due to reduced lipolysis and hence biohydrogenation through the combined effect of esterified C18:3 n-3 and seed protection. Additionally, C18:3 n-3 proportions were higher in rumen contents of RCS animals compared with WCS animals, which could be due to the activity of the polyphenol oxidase enzyme in the RC silages. Proportions of C18:3 n-3 were similar between treatments both for SC and intramuscular (IM) fat, whereas CLA c9t11 content was higher in the SC fat of BDS animals and lower in the IM fat of IRS animals compared with the other forage groups. No differences were found for C20:4 n-6, C20:5 n-3, C22:5 n-3 and C22:6 n-3 in the IM fat of the animals. Nevertheless, indices for desaturation and elongation activity in muscle of BDS animals suggest some stimulation of the first three steps of desaturation and elongation (Δ6-desaturase, elongase and Δ5-desaturase) of long-chain FA.     

Tissue fatty acid composition of pigs fed different fat sources

Dietary fat influences the physico-chemical properties of meat, and fatty acid (FA) composition may have implications on human health. The objectives of the experiment were to study tissue FA partitioning and the effect of dietary fat source on tissue FA composition. Seventy crossbred gilts (61.8 ± 5.2 kg BW average) were fed one of seven treatments: a diet containing a very low level of fat (no fat (NF)) and six fat-supplemented diets (10%: tallow (T), high-oleic sunflower oil (HOSF), sunflower oil (SFO), linseed oil (LO), fat blend (FB: 55% tallow, 35% SFO, 10% LO) and fish oil blend (FO: 40% fish oil, 60% LO). Differential tissue FA depositions were observed, with flare fat being the most saturated, followed by intermuscular, and subcutaneous being the least saturated. Monounsaturated fatty acid (MUFA) deposition showed an opposite tissue pattern. Subcutaneous fat showed the highest MUFAs, intermuscular fat showed intermediate values and flare fat showed the lowest MUFAs. Intramuscular polyunsaturated fatty acid (PUFA) content was less susceptible to dietary treatment modifications compared with other depots. Significant tissue FA modifications were observed due to dietary treatments, mainly in diets rich in PUFA. The saturated fatty acids (SFA) were high in NF-fed and low in HOSF-fed animals, MUFA were high in HOSF-fed and low in SFO-, LO- and FO-fed animals, while PUFA were high in SFO- and LO-fed and low in HOSF-, T- and NF-fed animals. Pigs fed LO and FB showed detectable levels of EPA, which depended on the linolenic content of the diet. The only effective way to increase tissue DHA contents was to add DHA in the diet through FO feeding. Araquidonic acid was high in SFO diets and low in LO and FB diets, and also high in intramuscular fat compared with other tissues. EPA and DHA were also high in intramuscular fat compared with other fat depots. The deposition of oleic and linoleic acids depended on the composition of dietary fat, as their deposition varied between diets, even at similar levels of intake of each FA. The NF diet resulted in the greatest proportion of SFAs (palmitic and stearic) of all treatments tested. SFAs were less susceptible to modification than MUFA in response to the different PUFA levels supplemented in the diet. T resulted in less fat deposition in some of the fat depots and more in others, suggesting that T could partition fat differently among fat depots.     

Effects of genomic selection for intramuscular fat content in breast muscle in Chinese local chickens

Improvements in living standards have resulted in consumers having higher expectations for chicken meat quality. This is particularly true in Asia, where there is high consumer preference for local breeds. Nothing is presently known about the effectiveness of using genomic selection (GS) strategies in chickens to genetically improve meat quality traits that cannot be measured in living potential parents. In this study, 724 Beijing‐You chickens were used as a training population; all were genotyped using Illumina 60K SNP chips, and intramuscular fat content in breast muscle (IMF_br) was measured. Birds in the GS line were selected based on genomic estimated breeding values, IMF_br being the sole trait. Genetic progress in one generation was compared to that from conventional family‐based selection, and both were evaluated against random‐bred controls. Results showed that relative to the random‐bred controls, IMF percentage was improved 9.62% using GS, comparable to the 10.38% improvement using family‐based selection. We quantified the effectiveness of GS when applied to a meat quality trait with low heritability in chickens. We plan to introduce custom SNP chips, appropriate for native chicken breeds in China, to assist in applying GS in local breeding and accelerate genetic gain.     

Reduced protein diets increase intramuscular fat of psoas major,a red muscle,in lean and fatty pig genotypes

The present study aims to assess the effects of pig’s genotype (lean v. fatty) and dietary protein level (control v. reduced) on intramuscular fat (IMF) content, fatty acid composition and fibre profile of psoas major, a representative red muscle in pig’s carcass scarcely studied relative to white longissimus lumborum. The experiment was conducted on 40 intact male pigs (20 Alentejana purebred and 20 Large White×Landrace×Pietrain crossbred) from 60 to 93 kg of live weight. Pigs were divided and allocated to four dietary groups: control protein diet equilibrated for lysine (17.5% of CP and 0.7% of lysine) and reduced protein diet (RPD) not equilibrated for lysine (13.1% of crude protein and 0.4% of lysine) within a 2×2 factorial arrangement (two genotypes and two diets). Alentejana purebred had higher IMF content (15.7%) and monounsaturated fatty acids (MUFA) (8.9%), whereas crossbred pigs had higher PM weight (46.3%) and polyunsaturated fatty acids (PUFA) (20.1%). The genotype also affected colour with higher lightness (15.1%) and yellowness (33.8%) and lower redness (9.9%) scores in crossbred pigs. In line with this, fatty pigs displayed more oxidative fibres (29.5%), whilst lean pigs had more glycolytic (54.4%). Relative to fatty acids, RPD increased MUFA (5.2%) and SFA (3.2%) but decreased PUFA (14.8%). Ultimately, RPD increased IMF content (15.7%) in the red muscle under study, with no impact on glycolytic to oxidative fibre type transformation.     

Daidzein enhances intramuscular fat deposition and improves meat quality in finishing steers

An experiment was conducted to determine the effects of soy isoflavone daidzein on carcass characteristics, fat deposition, meat quality, and blood metabolites in finishing steers. Fourteen crossbred steers were used in a 120-d finishing study. These steers were stratified by weight into groups and randomly allotted by group to one of two dietary treatments: (1) control and (2) daidzein (500 mg/kg concentrate). The steers were fed a 90% concentrate diet. Supplemental daidzein did not affect slaughter weight, hot carcass weight, and dressing percentage, but tended to reduce fat proportion (not including intramuscular fat) in carcass and backfat thickness of steers. The carcass bone proportion was greater in steers fed daidzein diets than those fed control diets. Daidzein supplementation reduced pH at 24 h after slaughtered and moisture content and increased isocitrate dehydrogenase activity, fat content (16.28% and 7.94%), marbling score (5.29 and 3.36), redness (a*), and chroma (C*) values in longissimus muscle relative to control treatment. The concentrations of blood metabolites including glucose, blood urea nitrogen, triglyceride, total cholesterol, non-esterified fatty acid, high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were all lower in steers fed daidzein diets than those fed control diets. Current results suggest that supplemental daidzein can affect lipid metabolism, increase intramuscular fat content and marbling score, and improve meat quality in finishing steers. Daidzein should be a promising feed additive for production of high-quality beef meat.     

Lipid composition of lactational diets influences the fatty acid profile of the progeny before and after suckling

The purpose of the present study was to compare the influence of adding no or 8% fat of varying sources (coconut oil, fish oil, rapeseed oil and sunflower oil) to diets for sows 1 week prior to farrowing and during lactation on the composition of fatty acids in plasma and tissues of the progeny while sucking and 3 weeks after weaning from the sow. A control diet without supplemental fat and four diets supplemented with 8% of coconut oil, rapeseed oil, fish oil or sunflower oil were provided to lactating sows (n = 15), and during the post-weaning period the same weaner diet was provided to all piglets (n = 15 litters), which were housed litterwise. The dietary ratio of n-6:n-3 fatty acids of the maternal diets largely influenced the progeny, as the ratio varying from 1.2 (fish oil) to 12.2 (sunflower oil) in the sow milk was reflected in plasma and adipose tissues of the sucking progeny. The liver showed similar variations according to dietary treatments, but a lower n-6:n-3 fatty acids ratio. From day 4 to later on during the suckling period, the concentration of C14:0, C16:0 and C18:1 in the liver of the piglets decreased, irrespective of the dietary treatments of sows. In plasma and liver, the total concentration of saturated fatty acids (SAFA), monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA) did not differ markedly in piglets sucking sows fed different dietary fatty acids, whereas the adipose tissue of piglets sucking sows fed sunflower oil and coconut oil showed the highest proportion of PUFA and SAFA, respectively. Weaning lowered the concentration of lipid-soluble extracts in plasma and the concentration of fatty acids in the liver of the piglets. Within the post-weaning period, dietary treatments of sows, rather than age of piglets, influenced the fatty acid composition of plasma and adipose tissue of the piglets, whereas the hepatic fatty acid profile was more affected by the age of the piglets during the post-weaning period. This study shows that the fatty acid profile of plasma and tissues of the progeny is highly dependent on the maternal dietary composition, and that the dietary impact persists for up to 3 weeks after the suckling period.     

Effect of oilseed type on milk fatty acid composition of individual cows,and also bulk tank milk fatty acid composition from commercial farms

Supplementing dairy cow diets with oilseed preparations has been shown to replace milk saturated fatty acids (SFA) with mono- and/or polyunsaturated fatty acids (MUFA, PUFA), which may reduce risk factors associated with cardio-metabolic diseases in humans consuming milk and dairy products. Previous studies demonstrating this are largely detailed, highly controlled experiments involving small numbers of animals, but in order to transfer this feeding strategy to commercial situations further studies are required involving whole herds varying in management practices. In experiment 1, three oilseed supplements (extruded linseed (EL), calcium salts of palm and linseed oil (CPLO) and milled rapeseed (MR)) were included in grass silage-based diets formulated to provide cows with ~350 g oil/day, and compared with a negative control (Control) diet containing no supplemental fat, and a positive control diet containing 350 g/cow per day oil as calcium salt of palm oil distillate (CPO). Diets were fed for 28-day periods in a 5×4 Latin Square design, and milk production, composition and fatty acid (FA) profile were analysed at the end of each period. Compared with Control, all lipid supplemented diets decreased milk fat SFA concentration by an average of 3.5 g/100 g FA, by replacement with both cis- and trans-MUFA/PUFA. Compared with CPO, only CPLO and MR resulted in lower milk SFA concentrations. In experiment 2, 24 commercial dairy farms (average herd size±SEM 191±19.3) from the south west of the United Kingdom were recruited and for a 1 month period asked to supplement their herd diets with either CPO, EL, CPLO or MR at the same inclusion level as the first study. Bulk tank milk was analysed weekly to determine FA concentration by Fourier Transform mid-IR spectroscopy prediction. After 4 weeks, EL, CPLO and MR all decreased herd milk SFA and increased MUFA to a similar extent (average −3.4 and +2.4 g/100 g FA, respectively) when compared with CPO. Differing responses observed between experiments 1 and 2 may be due in part to variations in farm management conditions (including basal diet) in experiment 2. This study demonstrates the importance of applying experimental research into commercial practice where variations in background conditions can augment different effects to those obtained under controlled conditions.     

Intramuscular fat in lamb muscle and the impact of selection for improved carcass lean meat yield

Intramuscular fat percentage (IMF%) has been shown to have a positive influence on the eating quality of red meat. Selection of Australian lambs for increased lean tissue and reduced carcass fatness using Australian Sheep Breeding Values has been shown to decrease IMF% of the Muscularis longissimus lumborum. The impact this selection has on the IMF% of other muscle depots is unknown. This study examined IMF% in five different muscles from 400 lambs (M. longissimus lumborum, Muscularis semimembranosus, Muscularis semitendinosus, Muscularis supraspinatus, Muscularis infraspinatus). The sires of these lambs had a broad range in carcass breeding values for post-weaning weight, eye muscle depth and fat depth over the 12^th rib (c-site fat depth). Results showed IMF% to be highest in the M. supraspinatus (4.87±0.1, P<0.01) and lowest in the M. semimembranosus (3.58±0.1, P<0.01). Hot carcass weight was positively associated with IMF% of all muscles. Selection for decreasing c-site fat depth reduced IMF% in the M. longissimus lumborum, M. semimembranosus and M. semitendinosus. Higher breeding values for post-weaning weight and eye muscle depth increased and decreased IMF%, respectively, but only in the lambs born as multiples and raised as singles. For each per cent increase in lean meat yield percentage (LMY%), there was a reduction in IMF% of 0.16 in all five muscles examined. Given the drive within the lamb industry to improve LMY%, our results indicate the importance of continued monitoring of IMF% throughout the different carcass regions, given its importance for eating quality.     

Effect of grazing pastures with different botanical composition by lambs on rumen fatty acid metabolism and fatty acid pattern of longissimus muscle and subcutaneous fat

In order to study the effect of grazing pastures with a different botanical composition on rumen and intramuscular fatty acid metabolism, 21 male lambs were assigned to three botanically different pastures: botanically diverse (BD) (consisting for 65% of a variety of grass species); Leguminosa rich (L) (consisting for 61% of Leguminosae) and intensive English ryegrass (IR) (with 69% Lolium perenne). Pastures were sampled weekly for 12 weeks for analysis of their fatty acid content and composition and on nine occasions to determine the botanical composition. Ruminal and abomasal contents were sampled at slaughter and muscle and subcutaneous fat 24 h after slaughter. All samples were prepared and analysed for fatty acid composition. The L pasture showed a higher fatty acid content (29.8 mg/g dry matter (DM) v. 18.5 and 25.5 mg/g DM, for BD and IR pastures, respectively), but the sum of the proportions of the major polyunsaturated fatty acids, C18:2 n-6 and C18:3 n-3, were similar for the three pastures (69.9, 69.4 and 71.1% of fatty acids methyl esters (FAME) for BD, L and IR pastures, respectively). The BD pasture was richer in C18:2 n-6 (18.2% of FAME), while IR pasture had a higher C18:3 n-3 content (57.2% of FAME). Rumen data showed that animals grazing the BD pasture presented higher proportions of biohydrogenation intermediates, mainly C18:1 t11, C18:2 t11c15 and CLA c9t11, suggesting an inhibition of biohydrogenation. These changes were associated with shifts in the rumen microbial population as indicated by differences in the rumen pattern of volatile fatty acids, microbial odd- and branched-chain fatty acids. In L pasture animals, the content of C18:2 n-6 and C18:3 n-3 in the abomasum and subcutaneous fat was higher. Finally, higher proportions of C20:4 n-6, C20:5 n-3 and C22:5 n-3 and higher indices for elongation and desaturation activity in the intramuscular fat of BD grazing animals suggest some stimulation of elongation and desaturation of long-chain fatty acids, although this also might have been provoked partially by reduced fat deposition (due to a lower growth rate of the animals).     

The effect of divergent selection for intramuscular fat on the domestic rabbit genome

An experiment of divergent selection for intramuscular fat was carried out at Universitat Politècnica de València. The high response of selection in intramuscular fat content, after nine generations of selection, and a multidimensional scaling analysis showed a high degree of genomic differentiation between the two divergent populations. Therefore, local genomic differences could link genomic regions, encompassing selective sweeps, to the trait used as selection criterion. In this sense, the aim of this study was to identify genomic regions related to intramuscular fat through three methods for detection of selection signatures and to generate a list of candidate genes. The methods implemented in this study were Wright’s fixation index, cross population composite likelihood ratio and cross population – extended haplotype homozygosity. Genomic data came from the 9th generation of the two populations divergently selected, 237 from Low line and 240 from High line. A high single nucleotide polymorphism (SNP) density array, Affymetrix Axiom OrcunSNP Array (around 200k SNPs), was used for genotyping samples. Several genomic regions distributed along rabbit chromosomes (OCU) were identified as signatures of selection (SNPs having a value above cut-off of 1%) within each method. In contrast, 8 genomic regions, harbouring 80 SNPs (OCU1, OCU3, OCU6, OCU7, OCU16 and OCU17), were identified by at least 2 methods and none by the 3 methods. In general, our results suggest that intramuscular fat selection influenced multiple genomic regions which can be a consequence of either only selection effect or the combined effect of selection and genetic drift. In addition, 73 genes were retrieved from the 8 selection signatures. After functional and enrichment analyses, the main genes into the selection signatures linked to energy, fatty acids, carbohydrates and lipid metabolic processes were ACER2, PLIN2, DENND4C, RPS6, RRAGA (OCU1), ST8SIA6, VIM (OCU16), RORA, GANC and PLA2G4B (OCU17). This genomic scan is the first study using rabbits from a divergent selection experiment. Our results pointed out a large polygenic component of the intramuscular fat content. Besides, promising positional candidate genes would be analysed in further studies in order to bear out their contributions to this trait and their feasible implications for rabbit breeding programmes.     

Effect of exogenous sterols on the growth and fatty acid composition of the oomycetePythium debaryanum

Exogenous ergosterol and cholesterol were found to affect the growth and lipogenesis of the oomycete fungusPythium debaryanum, which is unable to synthesize de novo steroid compounds. These sterols stimulated the growth of the fungus during its submerged cultivation in glucose-peptone medium. This was accompanied by the shortening of the lag phase, the lengthening of the period of active growth, and by a 3.7-or 4.3-fold increase in the maximum biomass in response to the addition of ergosterol or cholesterol, respectively. In the presence of ergosterol, the cellular content of polyenoic fatty acids increased, and the relative content of eicosapolyenoic fatty acids reached 31.4% of the total amount of fatty acids in cells. Conversely, cholesterol decreased the cellular content of polyenoic acids, and the relative content of eicosapolyenoic acids fell to 19.6% of the total amount of fatty acids. It may be inferred that exogenous sterols enhance the yield of pharmacologically active polyenoic acids because of the growth stimulation.     

High-throughput analysis of fatty acid composition of plasma glycerophospholipids

Plasma FA composition, a marker of FA status and dietary intake, is associated with health outcomes on a short- and long-term basis. Detailed investigation of the relationships between plasma FA composition and health requires the analysis of large numbers of samples, but manual sample preparation is very cumbersome and time consuming. We developed a high-throughput method for the analysis of FAs in plasma glycerophospholipids (GPs) with increased sensitivity. Sample preparation requires two simple steps: protein precipitation and subsequent base catalyzed methyl ester synthesis. Analysis of GP FAs is performed by gas chromatography. Coefficients of variation for FAs contributing more than 1% to total FAs are below 4%. Compared with the established reference method, results of the new method show good agreement and very good correlations (r > 0.9). The new method reduces the manual workload to about 10% of the reference method. Only 100 µl plasma volume is needed, which allows for the analysis of samples from infants. The method is well suitable for application in large clinical trials and epidemiological studies.     

Effect of high versus low doses of fat and vitamin A dietary supplementation on fatty acid composition of phospholipids in mice

Dietary fat and vitamin A provide important precursors for potent bioactive ligands of nuclear hormone receptors, which regulate various enzymes involved in lipid homeostasis, metabolism and inflammation. We determined the effects of dietary fat and dietary vitamin A on hepatic expression of two fatty acid metabolizing enzymes, elongase 6 (ELOVL6) and stearoyl-coenzyme A desaturase 1 (SCD1) and the concentration of saturated fatty acids (SAFA) and monounsaturated fatty acid (MUFA) of phospholipids in serum and liver. Mice (n = 6) were fed 4 weeks with diets containing 2, 5 and 25 % of fat or vitamin A (0, 2,500 and 326,500 RE/kg as retinyl palmitate). MUFAs and SAFAs were measured using GC and ESI–MS/MS. Hepatic expression of metabolizing enzymes was determined using QRT-PCR. ELOVL6 was significantly down-regulated in response to a high-fat diet (p < 0.001) and significantly up-regulated in response to low-fat diet (p < 0.05). SCD1 expression was significantly lower in high- versus low-fat diet (p < 0.05). The vitamin A content in the diet did not influence the hepatic expression of both enzymes. In plasma, the amounts of MUFAs bound to phospholipids significantly decreased in response to a high-fat diet and increased after a low-fat diet. This tendency was also observed in the liver for various phospholipids sub-classes. In summary, this study shows that fat content in the diet has a stronger impact than the content of vitamin A on hepatic gene expression of SCD1 and ELOVL6 and thereby on MUFA and SAFA concentrations in liver and plasma.     

Chemotaxonomy of planktonic cyanobacteria based on non-polar and 3-hydroxy fatty acid composition

Twenty-eight axenio planktonic cyanobacterial strains (10 Microcystis, three Oscillatoria, one Spirulina, one Aphanizomenon, 13 Anabaena) were investigated for their fatty acid composition by measurement of non-polar and hydroxy fatty acids. No 2-hydroxy fatty acids were detected in any strain, but 3-hydroxy fatty acids were detected in minor quantities in 24 strains. The highest portion of total fatty acids were non-polar fatty acids. Qualitative and quantitative analyses of 3-hydroxy fatty acids showed no taxonomic value in these strains, while the type of non-polar fatty acid composition was shown to be consistent within Microcystis and Anabaena strains, distinguishing them as type 4, characterized by the presence of 18:4, and type 2, characterized by 18:3 (α) of the Kenyon-Murata system. Two Oscillatoria agardhii Gomont strains were also included in the type 2 group due to the presence of 18: 3 (α), but the difference in characteristics of 16:2 and 16:3 between O. agardhii and Anabaena further divided type 2 into two subgroups: type 2A for Anabaena and type 2B for O. agardhii. A simplified unweighted pair group method with arithmetic averages (UPGMA) dendrogram demonstrated that the classification of 28 strains (Microcystis spp., Anabaena spp., Aphanizomenon flos-aquae (Lemmermann) Ralfs f. gracile (Lemmermann) Elenkin, O. agardhii and Spirullnasubsalsa Oersted ex Gomont based on numerical analysis of non-polar fatty acids corresponded to morphological species criteria, suggesting that non-polar fatty acid composition is a valuable chemical marker in the taxonomy of planktonic cyanobacteria. However, the fatty acid composition in Oscillatoria raciborskii is similar to that of Microcystis and very different from that of O. agardhii, suggesting its special position in Oscillatoria and the chemical diversity in the genus Oscillatoria.     

Neutral fat hydrolysis and long-chain fatty acid oxidation during anaerobic digestion of slaughterhouse wastewater

Neutral fat hydrolysis and long-chain fatty acid (LCFA) oxidation rates were determined during the digestion of slaughterhouse wastewater in anaerobic sequencing batch reactors operated at 25 degrees C. The experimental substrate consisted of filtered slaughterhouse wastewater supplemented with pork fat particles at various average initial sizes (D(in)) ranging from 60 to 450 microm. At the D(in) tested, there was no significant particle size effect on the first-order hydrolysis rate. The neutral fat hydrolysis rate averaged 0.63 +/- 0.07 d(-1). LCFA oxidation rate was modelled using a Monod-type equation. The maximum substrate utilization rate (kmax) and the half-saturation concentration (Ks) averaged 164 +/- 37 mg LCFA/L/d and 35 +/- 31 mg LCFA/L, respectively. Pork fat particle degradation was mainly controlled by LCFA oxidation rate and, to a lesser extent, by neutral fat hydrolysis rate. Hydrolysis pretreatment of fat-containing wastewaters and sludges should not substantially accelerate their anaerobic treatment. At a D(in) of 450 microm, fat particles were found to inhibit methane production during the initial 20 h of digestion. Inhibition of methane production in the early phase of digestion was the only significant effect of fat particle size on anaerobic digestion of pork slaughterhouse wastewater. Soluble COD could not be used to determine the rate of lipid hydrolysis due to LCFA adsorption on the biomass.