Feeding steers hay with extruded flaxseed together or sequentially has a profound effect on erythrocyte trans 11-18:1 (vaccenic acid)

Publication: Canadian Journal of Animal Science
19 April 2016


Extruded flaxseed and ground hay [25% and 75%; dry matter (DM) basis] were fed in a total mixed ration (TMR) or sequentially (non-TMR) to three pens of eight crossbred steers per diet. At 112 d, erythrocytes from non-TMR steers had 65% more vaccenic acid (trans 11-18:1) than TMR steers (P < 0.05).


De la graine de lin extrudée et du foin [25 % et 75 %; selon les matières sèches (DM — « dry matter »)] a été donné dans une ration totale mélangée (TMR — « total mixed ration ») ou séquentiellement (non TMR) à trois enclos de huit bouvillons croisés par diète. Au jour 112, les érythrocytes des bœufs non TMR contenaient 65 % plus d’acide vaccénique (trans 11-18:1) que les bouvillons TMR (P < 0,05). [Traduit par la Rédaction]
Several feeding studies have been conducted worldwide to increase the content of polyunsaturated fatty acid (PUFA) biohydrogenation products in beef and dairy products, particularly trans 11-18:1 [vaccenic acid (VA)] and cis 9, trans 11-18:2 [rumenic acid (RA)], due to their purported health benefits. Most RA in cattle, however, is synthesized endogenously from VA. Consequently, a strong relationship exists between the VA and RA contents of beef. High variability in VA between and within trials has been found in beef, and this includes analyses at the Lacombe Research and Development Centre for several studies where different ratios of forage and concentrate, with and without oils or oilseeds, were fed (Mapiye et al. 2013a, 2013c). Attempting to explain differences in results led us to examine two studies where similar flaxseed containing red clover silage-based diets were fed, but different levels of VA were found in beef (Mapiye et al. 2013a, 2013c). Anecdotal study observations suggested that in the study where high VA contents were found, there may have been some sorting of components within the total mixed ration (TMR), specifically preferential consumption of concentrate followed by forage. The objective of the present experiment was to feed steers, a flaxseed containing concentrate and hay in a TMR or sequentially (non-TMR) to determine if this would affect the VA content in erythrocytes. We hypothesized that if we fed TMR components sequentially in time, it would reduce biohydrogenation of fatty acids in the rumen and increase the percentage of VA in erythrocyte fatty acids. Blood is easily accessible during the course of a trial, and previous work has shown strong correlations between the VA content of erythrocytes and beef adipose tissue (Mapiye et al. 2013b).
For the present study, 48 British × Continental crossbred steers were used. Steers were part of a longer (240 d) feeding trial, but given the strength of 112-d erythrocyte results, independent reporting prior to the completion of the feeding trial was warranted. Results for the full trial, including carcass measurements, meat quality, sensory evaluation, and tissue fatty acid analysis will be reported elsewhere. Steers were weighed on two consecutive days to start the trial (325 ± 16 kg, mean ± SD) and stratified by weight to six pens of eight animals. Steers were then weighted every 28 d, and weights were used to calculate average daily gain (ADG). The diet was formulated to contain 75% tub ground grass/alfalfa hay and 25% linPRO™-R (a commercial concentrate containing flaxseed co-extruded with peas and alfalfa (50:44:6), O&T Farms, Regina, SK, Canada) on a DM basis. A premix was added to the linPRO™-R to provide 2200 IU vitamin A, 275 IU vitamin D, and 135 IU vitamin E per kg of diet DM. The diet was formulated to provide (% DM) 16.6 CP, 7.47 fat, 29.7 ADF, 1.26 Ca, 0.24 P, and 65% TDN. The steers were provided a free-choice trace mineral salt (96.5% NaCl, 4000 mg kg−1 Zn, 1600 mg kg−1 Fe, 1200 mg kg−1 Mn, 330 mg kg−1 Cu, 100 mg kg−1 I, 40 mg kg−1 Co). The major fatty acids (% of total fatty acids) in the diet were α-linolenic acid (40.8%), linoleic acid (19.2%), and oleic acid (8.86%). Three pens of eight steers were fed hay and linPRO™-R mixed together in a TMR, and three pens were fed hay and linPRO™-R sequentially (non-TMR). The TMR pens were fed daily at 0800. Enough TMR was provided to result in 10%–15% orts after 18 h and complete consumption by 24 h. The non-TMR pens were fed linPRO™-R daily at 0800, which was consumed within 1.5 h, and hay was fed 3–4 h later. The non-TMR ration was adjusted weekly to match the intake of the TMR. Feed provided to each pen was recorded daily and was then divided by the number of animals per pen (n = 8) to estimate individual feed intake (kg d−1). Feed samples were collected weekly and composited monthly prior to analyses. Animals were cared for in accordance with guidelines established by the Canadian Council on Animal Care (CCAC 2009).
On 0 and 112 d, blood was collected in the morning prior to feeding from the jugular vein using an 18-gauge needle into tubes containing EDTA anticoagulant (Vacutainer®, Becton, Dickinson and Company, Franklin Lakes, NJ, USA). After collection, the blood was centrifuged at 800g for 18 min, plasma and white cells were discarded, and erythrocytes were methylated according to Risé et al. (2005); fatty acid methyl esters (FAMEs) were extracted and stored in hexane at −20 °C until analyzed. FAMEs were analyzed by gas chromatography (GC) using a Bruker BR-2560 column (100 m, 25 μm ID, 0.2-μm film thickness) in a CP-3800 gas chromatograph equipped with an 8400-series autosampler (Varian Inc., Walnut Creek, CA, USA) using the 175 °C plateau temperature program and GC conditions described by Turner et al. (2015).
For Animal performance data [weights, dry matter intake (DMI), ADG, and feed efficiency], day 112 data were analyzed using the PROC MIXED procedure of SAS (SAS Institute Inc., Cary, NC, USA) with diet as a fixed effect and pen as the experimental unit. Erythrocyte VA data were subjected to repeated measures’ analysis using the PROC MIXED procedure of SAS, with day as the repeated factor and individual animals as the experimental unit. Using a first-order autoregressive covariance structure, the model incorporated the fixed effects of diet, day, diet × day. With the finding of a significant diet × time interaction, means were generated and separated using the LSMEANS and PDIFF options. Differences were declared significant at P < 0.05.
The DMI, ADG, and feed efficiency of steers fed the TMR and non-TMR diets that did not differ (P = 0.24–0.70; Table 1). When looking at erythrocyte fatty acid composition, a time by diet interaction was found for the percentage of VA in total fatty acids (P = 0.001; Fig. 1). The content of VA in erythrocyte fatty acids did not differ at 0 d (P = 0.42) but increased (P = 0.001) for both diets at 112 d, with the increase when feeding the non-TMR being 65% greater than the TMR (P = 0.001). Diet had significant effects on other trans 18:1 isomers analyzed, but concentrations for these averaged <0.5% of total fatty acids (data not shown). Based on the equation developed by (Mapiye et al. 2013b), the percentages of VA in the erythrocytes was predicted to generate perirenal fat VA concentrations of 2.2% on 0 d, and 8.4% for the TMR, and 13.2% for the non-TMR on 112 d. Previously, Mapiye et al. (2014) fed diets containing 15% rolled flaxseed in either a grass hay or red clover silage-based TMR and attained 4.7% VA in perirenal fat with no difference between diets. In a second trial, Mapiye et al. (2013c) fed 15% rolled flaxseed in a red clover silage-based TMR, where some sorting of ingredients occurred, and found 9.5% VA in perirenal fat. The high percentages of VA in erythrocyte fatty acids at 112 d of the present trial may have been achieved by feeding high-quality grass/alfalfa hay along with a more thoroughly processed flaxseed. The substantial increase in erythrocyte VA concentrations in the non-TMR group was related to the sequential consumption of the linPRO™-R followed by ground hay. This may have resulted in specific ruminal conditions such as increased rate of passage and reduced pH which have been shown to increase ruminal outflow of VA (Troegeler-Meynadier et al. 2003). In addition, feeding the diet components separately may have inhibited or reduced the number of ruminal bacteria involved in the last step in biohydrogenation from trans 18:1–18:0. Feeding fish oil (i.e., rich in docosahexaenoic acid and eicosapentaenoic acid) is known to inhibit the last step in C18-PUFA biohydrogenation from trans 18:1 to 18:0 (Shingfield et al. 2003) and, to our knowledge, this is the first report of a similar effect when feeding a concentrate enriched with 18:3n-3 (i.e., linPRO™-R) followed by grass/alfalfa hay. Results of the present trial suggest that the method of feeding management of supplementary sources of PUFA may influence the degree of deposition of PUFA biohydrogenation products in beef tissues.
Table 1.
Table 1. Steer weights and feeding performance when feed extruded flaxseed with hay together in a TMR or sequentially (non-TMR).
Fig. 1.
Fig. 1. Effects of feeding steers extruded flaxseed with hay together in a TMR or sequentially (non-TMR) on the percentage of trans 11-18:1 (vaccenic acid) in erythrocyte fatty acids. Means with different letters are different (P < 0.05). Error bars are + standard error of the mean.


This project was funded by the Alberta Meat and Livestock Agency (ALMA). The gift of linPRO™-R from O&T Farms (Regina, SK, Canada) is gratefully acknowledged. P. Vahmani gratefully acknowledges the receipt of an NSERC fellowship funded by an internal grant to MD through the Agriculture and Agri-Food Canada Peer Review Program.


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Published In

cover image Canadian Journal of Animal Science
Canadian Journal of Animal Science
Volume 96Number 3September 2016
Pages: 299 - 301
Editor: J. Plaizier


Received: 27 January 2016
Accepted: 22 March 2016
Accepted manuscript online: 18 April 2016
Version of record online: 19 April 2016


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Key Words

  1. flaxseed
  2. total mixed ration
  3. trans 11-18:1
  4. vaccenic acid


  1. lin/graine de lin
  2. ration totale mélangée
  3. trans 11-18:1
  4. acide vaccénique



P. Vahmani
Agriculture and Agri-Food Canada, Lacombe Research and Development Centre, 6000 C&E Trail, Lacombe, AB T4L 1W1, Canada.
D.C. Rolland
Agriculture and Agri-Food Canada, Lacombe Research and Development Centre, 6000 C&E Trail, Lacombe, AB T4L 1W1, Canada.
T.A. McAllister
Agriculture and Agri-Food Canada, Lethbridge Research and Development Centre, 1st Avenue South 5403, PO Box 3000, Lethbridge, AB T1J 4B1, Canada.
H.C. Block
Agriculture and Agri-Food Canada, Lacombe Research and Development Centre, 6000 C&E Trail, Lacombe, AB T4L 1W1, Canada.
S.D. Proctor
Metabolic and Cardiovascular Diseases Laboratory, Li Ka Shing Centre for Health Research Innovation, Alberta Diabetes and Mazankowski Institutes, University of Alberta, Edmonton, AB T6G 2E1, Canada.
L.L. Guan
Department of Agricultural Food and Nutritional Sciences, University of Alberta, Edmonton, AB T6G 2P5, Canada.
N. Prieto
Agriculture and Agri-Food Canada, Lacombe Research and Development Centre, 6000 C&E Trail, Lacombe, AB T4L 1W1, Canada.
J.L. Aalhus
Agriculture and Agri-Food Canada, Lacombe Research and Development Centre, 6000 C&E Trail, Lacombe, AB T4L 1W1, Canada.
M.E.R. Dugan [email protected]
Agriculture and Agri-Food Canada, Lacombe Research and Development Centre, 6000 C&E Trail, Lacombe, AB T4L 1W1, Canada.


Abbreviations: ADG, average daily gain; DM, dry matter; DMI, dry matter intake; FAMEs, fatty acid methyl esters; GC, gas chromatography; PUFA, polyunsaturated fatty acid; RA, rumenic acid; TMR, total mixed ration; VA, vaccenic acid.
© Her Majesty the Queen in right of Canada 2016. Permission for reuse (free in most cases) can be obtained from RightsLink.

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