Effects of sunflower oil infusions of Asparagopsis taxiformis on in vitro ruminal methane production and biohydrogenation of polyunsaturated fatty acids.

Asparagopsis taxiformis inhibits ruminal methane (CH4) production due to its bromoform (CHBr3) content. The immersion of A. taxiformis in edible vegetable oils allows the extraction and stabilization of the highly volatile CHBr3 in the oil phase. The objectives of this study were to explore the effects of adding sunflower oils with increasing concentrations of CHBr3 on in vitro ruminal methanogenesis and biohydrogenation. Five batches of 48-h in vitro incubations were performed in 14 fermentation bottles, using rumen inocula collected shortly after the slaughter of young crossbred bulls and 1 g of dry matter (DM) from a total diet of mixed feed without added oil (control) or with 60 µL of sunflower oil per gram of DM as the substrate. The treatments were the CHBr3 content in the oil added: 0 µg (B0), 25 µg (B25), 50 µg (B50), 75 µg (B75), 100 µg (B100), and 150 µg (B150) of CHBr3 per gram of substrate DM. Organic matter (OM) degradability, total gas, CH4, volatile fatty acids (VFA), long-chain fatty acids, and dimethyl acetals (DMA) were analyzed at the end of each incubation. Data were analyzed with a model considering the treatments as the fixed effect and the run as a random block and using orthogonal contrasts. Degradability of OM was higher in the control group and was unaffected by CHBr3 concentration. Total gas production per gram of degraded OM was unaffected by treatments and averaged 205 ± 29.8 mL/g. Methane (mL) production decreased linearly with increasing CHBr3 concentrations, with 33%, 47%, and 87% reductions for B75, B100, and B150, respectively. Total VFA concentration was unaffected by oil inclusion but was reduced by 20% in CHBr3-containing treatments, although without any dose-response pattern. The molar percentage of acetate decreased linearly, whereas propionate and butyrate increased linearly with the increasing CHBr3 dosage. Including oil in the diet decreased the branched-chain fatty acids and DMA content. Increasing CHBr3 concentrations did not affect branched-chain fatty acids, but linearly increased most of the identified DMA. Adding oil to the control diet increased the 18:2n-6, whereas increasing the concentration of CHBr3 had no effect on 18:2n-6 but decreased linearly the 18:0 and increased the trans-18:1 isomers. The results obtained provide evidence that oil immersions of A. taxiformis can successfully inhibit ruminal production of CH4 in vitro at doses of 100 and 150 µg/g DM, and simultaneously modulate biohydrogenation.

The effects of improving low dietary protein utilization on the proteome of lamb tissues.

Cistus ladanifer L. is a common shrub endemic to the Mediterranean region with high levels of condensed tannins (CT). CT form complexes with dietary protein resisting microbial degradation in the rumen, which enhances dietary protein utilization in ruminant diets. The objective of this study was to evaluate the utilization of CT in the diet of lambs on the proteomes of muscle, hepatic and adipose tissues. Twenty-four Merino Branco ram lambs were randomly allocated to three treatments (n = 8): C - control (160 g crude protein (CP)) per kg DM, RP - reduced protein (120 g CP/kg DM); and RPCT - reduced protein (120 g CP/kg DM) treated with CT extract. At the end of the trial, lambs were slaughtered and the longissimus lumborum muscle, hepatic and peri-renal adipose tissues sampled. A two-way approach was used for proteomic analysis: 2D-DIGE and nanoLC-MS. In the muscle, C lambs had lower abundance proteins that partake in the glycolysis pathway than the lambs of other treatments. Control lambs had lower abundance of Fe-carrying proteins in the hepatic tissue than RP and RPCT lambs. The latter lambs had highest abundance of hepatic flavin reductase. In the adipose tissue, C lambs had lowest abundance of fatty-acid synthase. SIGNIFICANCE: soybean meal is an expensive feedstuff in which intensive animal production systems heavily rely on. It is a source of protein extensively degraded in the rumen, leading to efficiency losses on dietary protein utilization during digestion. Protection of dietary protein from extensive ruminal degradation throughout the use of plants or extracts rich in CT allow an increase in the digestive utilization of feed proteins. In addition to enhance the protein digestive utilization, dietary CT may induce other beneficial effects in ruminants such as the improvement of the antioxidant status.

Effects of dietary inclusion of citrus pulp and rockrose soft stems and leaves on lamb meat quality and fatty acid composition.

Meat from lambs finished with high-starch diets often contains low concentration of vaccenic (t11-18:1) and rumenic (c9,t11-18:2) acids and high concentration of t10-18:1. We hypothesized that replacing cereals by dehydrated citrus pulp (DCP) and the inclusion of tanniferous feed sources in oil supplemented diets might reduce the accumulation of t10-18:1 and increase the t11-18:1 and c9,t11-18:2 in lamb meat, without affecting the productive performance. In total, 32 lambs were assigned to four diets which combine two factors: basal diet (BD) (cereals v. DCP) and Cistus ladanifer (CL) (0 v. 150 g/kg dry matter). Feed intake, average daily weight gain and carcass traits were not affected by treatments, except for dressing percentage that was reduced with DCP (P=0.046). Both DCP and C. ladanifer reduced tenderness and juiciness of meat, and C. ladanifer also reduced (P0.05) by diets. However, DCP increased the proportions of odd-chain FA (P=0.005) and several minor biohydrogenation (BH) intermediates in meat lipids. C. ladanifer had few effects on meat FA profile. The proportions of t11-18:1 and c9,t11-18:2 were high in all diets (5.4% and 1.5% of total FA, respectively) and were not influenced by the treatments. Basal diet and CL showed some significant interactions concerning FA composition of intramuscular fat. In diets without C. ladanifer, replacement of cereals by DCP increased the 18:0 (P<0.05) and decreased t10,c12-18:2 (P<0.05), t10-18:1 (P<0.10) and t10-/t11-18:1 ratio (P<0.10) with a large reduction of the individual variation for t10-18:1 and of t10-/t11-18:1 ratio. Combined with cereals, C. ladanifer increased 18:0 and reduced the BH intermediates in meat. Replacement of cereals by DCP seems to promote a more predictable FA profile in lamb meat, reducing the risk of t10-shifted BH pathways in the rumen.

Effect of feeding lambs with a tanniferous shrub (rockrose) and a vegetable oil blend on fatty acid composition of meat lipids.

The effects of feeding Cistus ladanifer (Cistus) and a blend of soybean and linseed oil (1 : 2 vol/vol) on fatty acid (FA) composition of lamb meat lipids and messenger RNA (mRNA) expression of desaturase enzymes was assessed. In total, 54 male lambs were randomly assigned to 18 pens and to nine diets, resulting from the combination of three inclusion levels of Cistus (50 v. 100 v. 200 g/kg of dry matter (DM)) and three inclusion levels of oil (0 v. 40 v. 80 g/kg of DM). The forage-to-concentrate ratio of the diets was 1 : 1. Longissimus muscle lipids were extracted, fractionated into neutral (NL) and polar lipid (PL) and FA methyl esters obtained and analyzed by GLC. The expression of genes encoding Δ5, Δ6 and Δ9 desaturases (fatty acid desaturase 1 (FADS1), fatty acid desaturase 2 (FADS2) and stearoyl CoA desaturase (SCD)) was determined. Intramuscular fat, NL and PL contents were not affected by oil or Cistus. Oil supplementation reduced (P<0.05) 16:0, c9-16:1, 17:0, c9-17:1 and c9-18:1 FA and increased (P<0.05) 18:2n-6, 18:3n-3 and the majority of biohydrogenation intermediates in NL. Cistus alone had few effects on FA of NL but interacted with oil (P<0.05) by increasing t10-18:1,t10,t12-18:2,t10,c12-18:2 and t7,c9-18:2. The t10-/t11-18:1 ratio increased with both Cistus and oil levels. The c9, t11-18:2 did not increase (P<0.05) with both oil and Cistus dietary inclusion. Oil reduced c9-16:1, 17:0, c9-17:1,c9-18:1, 20:4n-6, 22:4n-6 and 20:3n-9 proportions in PL, and increased 18:2n-6, 18:3n-3, 20:3n-3 and of most of the biohydrogenation intermediates. The Cistus had only minor effects on FA composition of PL. Cistus resulted in a reduction (P<0.05) of 20:5n-3 and 22:6n-3 in the meat PL. The expression level of SCD mRNA increased (P=0.015) with Cistus level, although a linear relationship with condensed tannins intake (P=0.11) could not be established. FADS1 mRNA expressed levels increased linearly (P=0.019) with condensed tannins intake. In summary, the inclusion of Cistus and oil in 1 : 1 forage-to-concentrate ratio diets resulted in a large increase in t10-18:1 and no increase in c9,t11-18:2 or n-3 long chain poor in polyunsaturated fatty acids in lamb meat.

Growth performance, carcass and meat quality of lambs supplemented with increasing levels of a tanniferous bush (Cistus ladanifer L.) and vegetable oils.

The effects of dietary inclusion of Cistus ladanifer L. (CL) and a vegetable oil blend were evaluated on growth performance,carcass and meat quality of fifty four lambs that were assigned to 9 diets, corresponding to 3 levels of CL(50, 100 and 200 g/kg DM) and 3 levels of oil inclusion (0, 40 and 80 g/kg DM). Treatments had no effects on growth rate. Oil depressed dry matter intake (P = 0.017), carcass muscle (P = 0.041) and increased (P = 0.016) kidney knob channel fat. Chemical and physical meat quality traits were not affected by treatments. Off-flavour perception was higher for 8% of oil (P b 0.001). The level of 100 g/kg DM of CL inclusion improved meat stability after 7 days of storage. Supplementation with linseed and soybean oils (2:1) was a good approach to improve meat nutritional value from feedlot lambs, increasing total n-3 PUFA.

The use of a tannin crude extract from Cistus ladanifer L. to protect soya-bean protein from degradation in the rumen.

Cistus ladanifer L. (CL) is a perennial shrub abundant in dry woods and dry land of Mediterranean zone, with high level of tannins. Tannins bind to protein, preventing its degradation in the digestive compartments. This tannin/protein complex may be advantageous when partially protecting good-quality feed protein from excessive rumen protein degradation. The objective of this trial was to use a CL phenol crude extract to prevent excessive rumen degradation of soya-bean meal protein. The phenolic compounds were extracted using an acetone/water solution (70:30, v/v). Soya-bean meal was then treated with this crude CL extract, containing 640 g of total phenols (TP) per kg of dry matter (DM), in order to obtain mixtures with 0, 12.5, 25, 50, 100 and 150 g of TP per kg DM. Three rumen-cannulated rams were used to assess in sacco rumen degradability of DM and nitrogen (N). The three-step in vitro procedure was used to determine intestinal digestibility. Increasing extract concentrations quadratically decreased the N-soluble fraction a (R2 = 0.96, P = 0.0001) and increased the non-soluble degradable fraction b (R2 = 0.92, P = 0.005). The rate of degradation c linearly decreased with CL extract doses (R2 = 0.44, P = 0.0065). For the effective rumen degradability of N, a linear reduction (R2 = 0.94, P < 0.0001) was observed. The in vitro intestinal digestibility of protein (ivID) quadratically decreased (R2 = 0.99, P < 0.0001) with TP inclusion and the rumen undegradable protein (RUP) showed a quadratic increase (R2 = 0.94, P = 0.0417). Total intestinal protein availability, computed from the RUP and ivID, linearly decreased with TP inclusion level (R2 = 0.45, P = 0.0033).