John R. Ashes

Incorporation of n−3 fatty acids of fish oil into tissue and serum lipids of ruminants

This study examines the biohydrogenation and utilization of the C20 and C22 polyenoic fatty acids in ruminants. Eicosapentaenoic (20∶5n−3) and docosahexaenoic (22∶6n−3) acids were not biohydrogenated to any significant extent by rumen microorganisms, whereas C18 polyenoic fatty acids were extensively hydrogenated. The feeding of protected fish oil increased the proportion of 20∶5 from 1% to 13–18% and 22∶6 from 2% to 7–9% in serum lipids and there were reductions in the proportion of stearic (18∶0) and linoleic (18∶2) acids. The proportion of 20∶5 in muscle phospholipids (PL) increased from 1.5% to 14.7% and 22∶6 from 1.0% to 4.2%; these acids were not incorporated into muscle or adipose tissue triacylglycerols (TAG). In the total PL of muscle, the incorporated 20∶5 and 22∶6 substituted primarily for oleic (18∶1) and/or linoleic (18∶2) acid, and there was no consistent change in the porportion of arachidonic (20∶4) acid.

Dietary n-3 and n-6 fatty acids alter avian metabolism: metabolism and abdominal fat deposition

The effects of dietary saturated fatty acids and polyunsaturated fatty acids (PUFA) of the n-3 and n-6 series on weight gain, body composition and substrate oxidation were investigated in broiler chickens. At 3 weeks of age three groups of chickens (n 30; ten birds per group) were fed the fat-enriched experimental diets for 5 weeks. These diets were isonitrogenous, isoenergetic and contained 208 g protein/kg and 80 g edible tallow, fish oil or sunflower oil/kg; the dietary fatty acid profiles were thus dominated by saturated fatty acids, n-3 PUFA or n-6 PUFA respectively. Resting RQ was measured in five birds from each treatment group during weeks 4 and 5 of the experiment. There were no significant differences between treatments in total feed intake or final body mass. Birds fed the PUFA diets had lower RQ and significantly reduced abdominal fat pad weights (P<0.01) compared with those fed tallow. The dietary lipid profile changes resulted in significantly greater partitioning of energy into lean tissue than into fat tissue (calculated as breast lean tissue weight:abdominal fat mass) in the PUFA groups compared with the saturated fat group (P<0.01; with no difference between the n-3 and n-6 PUFA groups). In addition, the PUFA-rich diets lowered plasma concentrations of serum triacylglycerols and cholesterol. The findings indicate that dietary fatty acid profile influences nutrient partitioning in broiler chickens.

Utilisation of fish oil in ruminants

Abstract A study was conducted to determine the transfer of eicosapentaenoic acid (EPA, 20:5) and docosahexaenoic acid (DHA, 22:6) from fish oil into goats’ milk. Goats were sequentially offered three diets: control (C) pellets (lucerne hay-oat grain: 60/40 w/w), C plus tuna oil protected against ruminal biohydrogenation (PTO pellets), and C plus unprotected tuna oil (UTO pellets). In supplemented diets, tuna oil constituted 3% of total dry matter (DM), and each supplement was fed for 7 days, with 12 days allowed between the two fish oil feeding periods to minimise carry-over effects. Dry matter intake, milk yield, protein and fat yield were reduced by feeding UTO, but not PTO, pellets. Goats produced ω-3 enriched milk (0.3–0.5% EPA and 1.01–1.12% DHA) when fed either supplement. The rate of transfer of dietary EPA and DHA to milk ranged from 3.5 to 7.6%. Significant transfer of EPA and DHA from tuna oil into goat milk, without deleterious effects on intake or milk yield is possible, provided that the oil supplement is substantially protected against ruminal biohydrogenation.

Protection of conjugated linoleic acids from ruminal hydrogenation and their incorporation into milk fat

In vitro incubations were used to assess the hydrogenation of conjugated linoleic acid (CLA) isomers 9-cis 11-trans and 10-trans 12-cis present in synthetically produced CLA-60. About 80‐ 90% of the unprotected CLA was hydrogenated when incubated at 388C for 24 h anaerobically with sheep rumen fluid, the main end product of hydrogenation being trans-octadecaenoic acid (C18:1). Encapsulation of the CLA in a matrix of protein provided a protection of about 70% with a 30% hydrogenation of the CLA isomers, resulting in no significant change in the trans-C18:1 but an increase in the level of stearic acid (C18:0). Feeding sheep with unprotected CLA or protected CLA increased the proportion of isomers 9-cis 11-trans and 10-trans 12-cis in abomasal digesta. The concentration of the CLA isomers leaving the abomasum and available for absorption at the small intestine was about 3.5‐4% higher for the protected CLA, confirming protection imparted by encapsulation. Feeding lactating goats with protected CLA increased the proportion of isomers 9-cis 11-trans and 10-trans 12-cis in milk fat. The total CLA levels were enhanced by about 10-fold above the control levels present in milk fat with an efficiency of transfer into milk fat of 36‐41% and 21‐30%, respectively, for the two isomers. # 2000 Published by Elsevier Science B.V.

In-vitro assessment of fat supplements for ruminants

Abstract In-vitro anaerobic incubations were used to assess the degree of metabolism or rumen inertness of different fat supplements containing either triacyglycerols and/or free fatty acids. The extent of triacylglycerol lipolysis and biohydrogenation of the unsaturated fatty acids, oleic acid (C18:1) and linoleic acid (C18:2) was monitored after 24h incubation in rumen fluid. These studies indicate that the degree of metabolism of different fat sources varies from 15 to 95%. For untreated oil, extruded oil-seed and pelleted (prilled) fat supplements the degradation values were 95, 70, and 65% respectively. The hydrogenation of calcium salts of C18 monounsaturated fatty acids when mixed with predominately C16 and C18 saturated fatty acids was 45%. The lowest degree of metabolism (

Effect of feeding different fat supplements on the fatty acid composition of goat milk

The effect of feeding dietary fat supplements on the fatty acid composition of goat milk was examined. Inclusion of canola and soybean (8020; ww) oilseed supplement protected from ruminal hydrogenation, significantly increased the proportion of C18:1 (oleic acid), C18:2 (linoleic acid) and decreased the proportion of C16:0 (palmitic acid) and C14:0 (myristic acid), while there was a small increase in C18:0 (stearic acid). Feeding protected cotton seed significantly increased C18:2 and C18:0, but there was a reduction in C18:1 while the C16:0 was unchanged. When combinations of protected cotton seed and protected-canola soybean (8020; ww) were fed, a level of 20–25% incorporation of protected cotton seed was sufficient to inhibit the desaturase enzyme, with an increase in the proportion of C18:0. In contrast, feeding calcium salts of fatty acids, a predominantly saturated fatty acid supplement, increased C16:0 and reduced C10:0 (decanoic acid) and C14:0. Feeding fat supplements of different fatty acid compositions and varying levels of inertness in the rumen will enable significant manipulation of the fatty acid composition of milk fat.

Dietary n-3 and n-6 fatty acids alter avian metabolism: molecular-species composition of breast-muscle phospholipids

The effects of diets high in n-3 polyunsaturated fatty acids (PUFA; provided by fish oil), n-6 PUFA (sunflower oil) or in more-saturated fatty acids (tallow) on the distribution of subclasses of choline phospholipids (PC) and ethanolamine phospholipids (PE) from the breast muscle of broiler chickens were examined. Supplementation with the different fatty acids had no effect on the distribution of phospholipid subclasses. Feeding sunflower oil or tallow gave a molecular-species profile similar in both fatty acid subtype and proportion. In the diacyl PC phospholipids, 16 : 0-18 : 1n-9 and 16 : 0-18 : 2n-6 accounted for approximately 60 % of the total molecular species, whereas for the alkylenyl PC the predominant species were 16 : 0-18 : 1n-9 and 16 : 0-20 : 4n-6. Of the diacyl PE the dominant species was 18 : 0-20 : 4n-6 which accounted for 50 % of the molecular species, and of the alkylenyl PE the dominant species were 16 : 0-18 : 1n-9, 16 : 0-20 : 4n-6 and 18 : 0-20 : 4n-6. Supplementation with fish oil significantly increased levels of both eicosapentaenoic acid (20 : 5n-3) and docosahexaenoic acid (22 : 6n-3) in PC and PE when compared with either sunflower oil or tallow supplementation. The increase in the n-3 PUFA incorporation was associated with a corresponding decrease in the proportion of arachidonic acid (20 : 4n-6) in both PC and PE. Different dietary fats induce different patterns of fatty acid incorporation and substitution in the sn-2 position of the diacyl and alkylenyl PC and PE of avian breast muscle, and this finding is indicative of selective acyl remodelling in these two phospholipids.

Increased efficiency of wool growth and live weight gain in Merino sheep fed transgenic lupin seed containing sunflower albumin

The aim of this experiment was to assess, using sheep, the nutritive value of lupin seed transgenically modified to contain sunflower seed albumin. Eighty Merino wether sheep of mean live weight 32.3 kg were divided into two groups and fed 796 g dry matter (DM) day−1 of a cereal hay-based diet containing 350 g kg−1 of either the transgenic or parent (unmodified) lupin seed for 6 weeks. Measurements were made of wool growth and live weight gain. After 6 weeks, half the sheep in each group were selected for a urine and faeces balance study in which organic matter (OM), nitrogen (N) and urinary purine metabolites were measured. Blood samples were taken from all sheep at the beginning and end of treatment and analysed for amino acids and plasma metabolites. A comprehensive chemical analysis of the grains showed that there was little difference between them in terms of most nutritional components, but the transgenic lupin seed contained a 2.3-fold higher methionine concentration and 1.3-fold higher cysteine than did the parent. There were no significant differences between grains in OM digestibility, rumen microbial protein synthesis or in sacco degradability of dry matter. Sheep fed the transgenic lupin grain had an 8% higher rate of wool growth (P   0.1). Plasma urea N was lower in the sheep fed the transgenic grain than those fed the parent grain (6.5 vs 6.8 mmol l−1, P < 0.05). The results show that genetic modification of a feed grain can improve its nutritive value for ruminants. The size and nature of the responses were consistent with the transgenic lupins providing more methionine to the tissues, a first-limiting amino acid for sheep. © 2000 Society of Chemical Industry

Effect of particle size and lipid composition of bovine blood high density lipoprotein on its function as a carrier of β-carotene

As part of a study on the influence of dietary lipids on vitamin transport and metabolism in lactatinf cows, we have examined the β-carotene content and other properties of fractions of the high-density lipoprotein (HDL, density 1.05–1.16 g/ml) of bovine blood. Our purpose was primarily to explain previous results indicating that feeding cows polyunsaturated lipids alters the properties of the HDL and increases the concentration of β-carotene in the blood but not in the milk. Fractions of HDL of different particle size were prepared by gel-filtration chromatography and the particle diameters measured by electron microscopy. We found that large HDL particles contain more β-carotene per unit weight than small particles. Furthermore the HDL from cows fed lipid-rich diets with a high proportion of linoleic-acid residues, which had been protected against microbial degradation in the rumen, had a high percentage of HDL particles with large diameters. The blood from these cows had a higher concentration of β-carotene than before feeding polyunsaturated lipids, but their milk had a lower concentration. We suggest that HDL is the main store of β-carotene in bovine blood. Moreover the concentration of β-carotene in blood is increased by feeding polyunsaturated lipids largely because of the increase in the percentage of large HDL particles, which contain more β-carotene. The effect on the concentration of β-carotene in milk implies that the transfer mechanism is less efficient as a result of feeding polyunsaturated lipids. This lower efficiency may be due in part to the higher percentage of large HDL particles.

Nutritional availability of amino acids from protein cross-linked to protect against degradation in the rumen

Casein was labelled with pairs of radioactive amino acids, lysine, tyrosine and leucine, one with 14C and the other with 3H, by jugular infusion into lactating goats followed by isolation of the double-labelled casein from the milk. Total milk protein was similarly labelled by jugular infusion of [35S]cystine. U-14C-labelled fraction-1 leaf protein was isolated from lucerne (Medicago sativa) grown in an atmosphere of 14CO2. The proteins were treated with different levels (333 and 667 mmol/kg protein) of formaldehyde, glutaraldehyde and glyoxal. Absorption from the small intestine was measured in sheep with fistulas in the abomasum and terminal ileum, using Cr-EDTA as the digesta flow marker, by introducing radioactive casein into the abomasum. Lysine, tyrosine and cystine became increasingly unavailable for absorption from the small intestine of sheep with increasing levels of aldehyde. At the lower level (333 mmol/kg) the proportions of the amino acids that were unavailable were 0.192, 0.051 and 0.123 respectively. At the higher level of formaldehyde (667 mmol/kg) the corresponding values were 0.335, 0.201 and 0.432 respectively. Leucine was not made unavailable with formaldehyde. The proportions of lysine, tyrosine and leucine that were unavailable were higher, on a molar basis, after treatment of the proteins with the dialdehydes glutaraldehyde and glyoxal than after treatment with formaldehyde. However, the extent of protein protection provided by the dialdehydes in the rumen, measured using an in vitro procedure, was lower.