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The red seaweed Asparagopsis taxiformis has a potent antimethanogenic effect, which has been proven both in vitro and in vivo. Vegetable oil immersions of this seaweed (hereafter Bromoil) help stabilise the bromoform (CHBr 3 ) responsible for its antimethanogenic effect. We evaluate the effects of increasing the levels of CHBr 3 in lamb diets on growth performance, methane (CH 4 ) production, animal health and meat quality. Twenty-four Merino Branco ram lambs were fed a ground complete compound feed, supplemented with 50 mL/kg DM of sunflower oil with different CHBr 3 content. The treatments were defined by the CHBr 3 doses in the oil: 0 mg (control - B0), 15 mg (B15), 30 mg (B30) and 45 mg (B45) of CHBr 3 per kg of feed DM. The feed was prepared daily by mixing Bromoil with the compound feed. At the end of the experiment, the lambs were sacrificed, the ruminal content was collected for in vitro fermentation to evaluate CH 4 production and organic matter (OM) degradability, and the rumen mucosa was sampled for histological examination. Meat samples were collected for chemical composition and CHBr 3 analysis. The half-life of CHBr 3 in the air-exposed feed was 3.98 h making it very difficult to establish the practiced level of CHBr 3 supplementation. Lambs-fed treatments B30 and B45 decreased DM intake by up to 28%. Average daily gain was also reduced due to CHBr 3 supplementation, with B45 showing results 40% lower than B0. DM feed conversion ratio was similar for all treatments. The degradability of OM, the volume of total gas and of gas without CH 4 were unaffected by the experimental treatments, evaluated by the in vitro method. However, the volume of CH 4 decreased by up to 75% for treatments above 30 mg/kg DM, while the yield of CH 4 /g OM degraded was reduced by up to 78% with treatments above 30 mg/kg DM. Meat chemical composition was not affected by Bromoil supplementation and no traces of CHBr 3 were found in meat samples. During this experiment, the animals presented normal health and behaviour. However, postslaughter examination of the rumen showed distinct lesions on the ventral region of the rumen mucosa of animals supplemented with Bromoil. These lesions were more severe in the animals receiving treatments B30 and B45. This research determined that although concentrations of CHBr 3 in the diet above 30 mg/kg DM helped to reduce CH 4 emissions, it negatively affected the performance and rumen wall., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Asparagopsis taxiformis inhibits ruminal methane (CH 4 ) production due to its bromoform (CHBr 3 ) content. The immersion of A. taxiformis in edible vegetable oils allows the extraction and stabilization of the highly volatile CHBr 3 in the oil phase. The objectives of this study were to explore the effects of adding sunflower oils with increasing concentrations of CHBr 3 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 CHBr 3 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 CHBr 3 per gram of substrate DM. Organic matter (OM) degradability, total gas, CH 4 , 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 CHBr 3 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 CHBr 3 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 CHBr 3 -containing treatments, although without any dose-response pattern. The molar percentage of acetate decreased linearly, whereas propionate and butyrate increased linearly with the increasing CHBr 3 dosage. Including oil in the diet decreased the branched-chain fatty acids and DMA content. Increasing CHBr 3 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 CHBr 3 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 CH 4 in vitro at doses of 100 and 150 μg/g DM, and simultaneously modulate biohydrogenation., (The Authors. Published by Elsevier Inc. and Fass Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

Two groups of 8 individually housed young crossbred-bulls, in the finishing period, were used to test the effect of a Total Mixed Ration diet with high forage content (54% DM), low starch content (14% DM), supplemented with sunflower seeds (10% DM) (HFS) on growth performance, carcass and meat quality, fatty acid profile and carbon footprint, with reference to a conventional concentrate-based (90% DM) (Control) diet. The experiment lasted 64 days before slaughter. During the experiment, feed intake was monitored daily and live weight every 14 days. Individual CH 4 emissions were assessed at 16-days intervals, using a GreenFeed for Large Animal unit. Feed intake and feed conversion ratio were higher for HFS diet, but average daily weight gain and feeding costs were similar for the two diets. Dressing percentage was reduced with HFS diet. The HFS increased redness, yellowness and Chroma of subcutaneous fat, but did not compromise commercial value of the carcasses. Meat colour, shear force, or sensory parameters were not affected by diet. The HFS diet allowed a healthier FA profile, due to the higher proportions of 18:3n-3, t11-18:1 and c9,t11-18:2 and the lower proportion of t10-18:1. The HFS diet did not reduce the carbon footprint in the finishing period of young bulls, due to increased digestive CH 4 emissions. The results of this experiment showed that the HFS diet can be an alternative to the conventional diets used in finishing young-bulls. Although it may result in a slight reduction in animal performance, it has a strong impact on reducing dependence on inputs from outside the farm., Competing Interests: Declaration of Competing Interest None., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Thirty-two bulls were assigned to four total mixed biodiverse haylage-based diets to evaluate the effects of partial substitution of grains by agroindustrial byproducts, sunflower seeds (SS) supplementation and haylage level on growth, in vitro methane production and carcass and meat quality. Dietary treatments included a grain-based diet with 30% grain and haylage:concentrate ratio (H:C) of 60:40 (DM basis) (MCe); a by-product-based diet where 50% of the grain was substituted for by-products (H:C, 60:40) (MBp); a byproducts diet with 10% sunflower seed and 90% (DM) MBp (H:C, 54:46) (MBpSS); and a byproducts, SS diet with increased haylage (H:C, 67.5:32.5) (HBpSS). Dry matter intake and growth rate were lower in HBpSS, but feed conversion ratio was unaffected by diet. In vitro methane emissions were reduced by SS. Meat colour and shear force were similar among diets. Lipid oxidation in cooked meat was reduced and fatty acid composition was improved with SS. Biodiverse haylage-based diets may be a viable option for finishing bulls., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

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.

Background: Tannins are polyphenolic compounds able to modify the ruminal biohydrogenation (BH) of unsaturated fatty acids, but their activity may vary among different tannin sources. The effect of rockrose (Cistus ladanifer) on BH has never been compared with other more common tannin sources. Tannin extracts (100 g kg -1 substrate dry matter) from chestnut (CH), quebracho (QB), grape seed (GS) and rockrose (CL) were incubated in vitro for 6 h with ruminal fluid using as substrate a feed containing 60 g kg -1 of sunflower oil. A control treatment with no added tannins was also included., Results: Compared with the control, GS and CL, but not CH and QB, increased (P < 0.05) the disappearance of c9,c12-18:2 with a consequent larger production of c9,t11-18:2 and t11-18:1. However, no differences among treatments (P > 0.05) were observed for the disappearance of c9-18:1 and c9,c12,c15-18:3. The production of 18:0 was not different (P > 0.05) among treatments, although its proportion in the total BH products was lower (P < 0.05) for GS than for the other treatments., Conclusion: Condensed tannins from GS and, to a lesser extent, from CL stimulate the first steps of BH, without a clear inhibition of 18:0 production. © 2016 Society of Chemical Industry., (© 2016 Society of Chemical Industry.)

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.

This study evaluates the effect of the replacement of cereals by dried citrus pulp (DCP) in diets supplemented with 5% of soybean oil, on ewe milk yield and composition, including milk fatty acid (FA). Four Serra da Estrela multiparous ewes in the second month of lactation were used in a double 2×2 Latin square design. Ewes were individually penned and milked twice a day with an 8-h interval. Each experimental period included 14 d of diet adaptation followed by 5d of measurements and sampling. The 2 diets included on dry matter basis 450 g/kg of corn silage and 550 g/kg of either a soybean oil-supplemented concentrate meal containing barley and maize (cereal) or dried citrus pulp (DCP; citrus). Feed was offered ad libitum, considering 10% of orts, and intake was measured daily. Milk yield was higher and dry matter intake tended to be higher with the citrus diet. Milk composition and technological properties for cheese production were not affected by treatments, except for lactose, which was lower with the citrus diet. Replacement of cereals by DCP resulted in a 3-percentage-point decrease of both 18:0 and cis-9-18:1 that were mostly compensated by the 4.19- and 1.68-percentage-point increases of trans-11-18:1 and cis-9,trans-11-18:2, respectively. The intake of C18 FA tended to increase with the citrus diet compared with the cereal diet, but the apparent transfer of 18:2n-6 and of 18:3n-3 did not differ between diets. The milk output of C18 FA increased with the citrus compared with the cereal diet, mostly due to the increase of trans-11-18:1 and cis-9,trans-11-18:2 because the daily milk output of 18:0, trans-10-18:1, cis-9-18:1, 18:2n-6 and 18:3n-3 did not differ between diets. Replacing cereals with DCP in an oil-supplemented diet resulted in a selective increase of trans-11-18:1 and cis-9,trans-11-18:2 in milk, with no major effect on other biohydrogenation intermediates., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

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.

Thirty-six Merino Branco lambs were assigned to six dietary treatments: control diet (C) consisting of 90% dehydrated lucerne and 10% wheat bran; C with 6% of oil blend (CO); C with 2.5% of grape seed extract (GS); GS with 6% of oil blend (GSO); C with 25% of Cistus ladanifer (CL), and CL with 6% of oil blend (CLO). Meat lipid and colour stability was then evaluated during 7 days of storage. The effect of inclusion of grape seed extract and C. ladanifer in diets on meat sensory properties was also evaluated. Meat antioxidant potential, determined after oxidation induction by a ferrous/hydrogen peroxide system, decreased with oil supplementation (P<0.001), but inclusion of grape seed extract and C. ladanifer in diets protected the meat against lipid oxidation (P=0.036). Meat colour was not affected by diets. Inclusion of grape seed extract and C. ladanifer in diets did not change the sensory properties of meat., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Thirty-six lambs were used in a 6 week experiment to evaluate the effect of vegetable oil blend supplementation (0 vs 60 g/kg of dry matter (DM)) and two dietary condensed tannin sources, grape seed extract (0 vs 25 g/kg of DM) and Cistus ladanifer L. (0 vs 250 g/kg of DM), on fatty acid (FA) composition of abomasal digesta and intramuscular polar and neutral lipids. Grape seed extract did not affect the FA profile of abomasal digesta or muscle lipid fractions. C. ladanifer had a minor effect in lambs fed diets with no oil but greatly changed the abomasal and muscle FA profiles in oil-supplemented lambs. It decreased 18:0 and increased 18:1 trans-11 in abomasal digesta and increased 18:1 trans-11 and 18:2 cis-9,trans-11 (P = 0.062) in muscle neutral lipids, resulting in an important enrichment of meat 18:2 cis-9,trans-11 when compared to other oil-supplemented diets (19.2 vs 41.7 mg/100 g of muscle).

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).