Effects of replacing canola meal with camelina expeller on intake, total tract digestibility, and feeding behavior of beef heifers fed high-concentrate diets Hèctor Salas,† Lorena Castillejos,†,1, Cristian Faturi,‡ and Alfred Ferret† Animal Nutrition and Welfare Service, Department of Animal and Food Sciences, Universitat Autònoma de † Barcelona, 08193 Bellaterra, Spain; and ‡Instituto da Saúde e Produçao Animal, Universidade Federal Rural da Amazônia, 66.077-830 Belém, Pará, Brazil ABSTRACT: To assess the effect of inclusion (38.6 vs. 8.7 kg/d; P < 0.001). When molasses of camelina expeller in beef cattle diets, 24 were added to the diet, the preference for CM Simmental heifers were used. Two experiments was maintained (39.1 vs. 9.8 kg/d; P < 0.001). were carried out. In the first, two free-choice Dry matter (DM), organic matter (OM), crude tests, one without and another with molasses, protein (CP), and neutral detergent fiber (NDF) were conducted to know the preference of ani- intake was unaffected by the level of replacement mals for a total mixed ration (TMR) made with a of CM by CE (P > 0.10), and there was no effect 90 to 10 concentrate to barley straw ratio, where of this replacement on DM, OM, CP, and NDF canola meal (CM) or camelina expeller (CE) apparent digestibility (P > 0.10). Intake of long was used in the concentrate as a protein source. particle size increased lineally as CE proportion Heifers were allotted in four pens with two inde- increased (P = 0.015). In addition, extension of pendent feedbunks, one for each diet. In the sec- sorting behavior for long particle size tended to ond, a replicated 4 × 4 Latin square design was increase lineally (P = 0.07), and sorting against used to assess the effects of the replacement of this particle size was detected in 0CE and 3CE, CM with CE on intake, digestibility, and sorting but not in 6CE and 9CE (P < 0.05). However, the and animal behavior. The experiment was per- results recorded for long particle size intake and formed in four 28-d periods during which groups for sorting behavior against these particles did of three animals were allotted in each pen of 12.5 not translate into more time spent ruminating in m2. Diets were formulated with a 90 to 10 con- heifers fed diets with higher proportion of CE. In centrate to barley straw ratio and fed as TMR, conclusion, when canola meal was replaced with and they were designed to contain 1) CM as main camelina expeller at more than 14% of inclusion, protein source and 0% of CE (0CE), 2) 3% of CE heifers preferred the canola meal diet. However, replacing CM (3CE), 3) 6% of CE replacing CM replacing canola meal with camelina expeller up (6CE), and 4) 9% of CE replacing CM (9CE). to 9% of inclusion in diets for beef cattle did not In the free-choice test without molasses, heifers affect intake and digestibility but promoted a showed a greater preference for CM than for CE greater intake of long particle size of barley straw. Key words: beef cattle, camelina expeller, canola meal © The Author(s) 2020. Published by Oxford University Press on behalf of the American Society of Animal Science. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribu- tion, and reproduction in any medium, provided the original work is properly cited. Transl. Anim. Sci. 2020.4:922–932 doi: 10.1093/tas/txaa050 Corresponding author: Lorena.Castillejos@uab.cat 1 Received January 31, 2020. Accepted April 23, 2020. 922 Camelina expeller in feedlot diets 923 INTRODUCTION The main concern with feeding camelina coproducts is the antinutritional compounds found Common plant protein sources, such as soybean in all Brassica species (Tripathi and Mishra, 2007). meal and canola meal, are expensive and subject to Camelina contains glucosinolates and erucic acid volatile prices. These economic circumstances are (Zubr, 1997). Glucosinolates can induce a lower ac- driving the research of potential new protein sources tivity of the thyroid gland and cause metabolism for beef cattle diets that can reduce ration cost with- disturbances, and erucic acid induces myocardial out compromising animal performance. Camelina lipidosis in rats (Putnam et al., 1993). Glucosinolate sativa is a plant with low agronomic requirements and content is higher in camelina than in canola meal very tolerant to adverse climatic conditions (Putnam (20 to 30 mmol/g vs. <10 mmol/g, respectively). et al., 1993). The industry’s interest in using camelina However, camelina does not contain the progoitrin seeds to produce biofuel has generated coproducts that forms the toxic goitrin, which can decrease that can be useful as protein sources to feed animals. hormone production in the thyroid, so camelina Colombini et al. (2012) studied different genotypes coproducts are of moderate toxicity with regard to of Camelina sativa and, after biodiesel extraction, their glucosinolate content (Matthäus, 1997). The stated that they have potential for use in ruminant content of erucic acid is usually higher in camelina rations as a high-quality protein source. Basically, coproducts than in canola meal (2% to 5% vs. <2%, two coproducts can be considered: camelina expeller respectively), but much lower than in high-erucic after a mechanical oil extraction, and camelina meal rapeseed oil. Our hypothesis was that considering when, in addition to mechanical extraction, a solvent the lower price of camelina expeller in comparison is also involved, raising the price of camelina meal with camelina meal, the potential nutritive value with regard to camelina expeller. of camelina expeller and the similarity of chemical Camelina coproducts have been used in cattle characteristics and efficiency of microbial protein diets for growing heifers (Moriel et al., 2011; synthesis with canola meal, camelina expeller could Lawrence et al., 2016), dairy cows (Hurtaud and be considered a protein source in beef cattle diets. Peyraud, 2007; Halmemies-Beauchet-Filleau et al., Thus, due to the lack of information on the effects 2011), and beef cattle (Cappellozza et al., 2012). of inclusion of camelina expeller on high-concen- Some of these experiments did not detect effects on trate diets for beef cattle, our objective was to study intake or animal performance (Moriel et al., 2011; its effects on intake, nutrient digestibility, sorting, Halmemies-Beauchet-Filleau et al., 2011; Lawrence and feeding behavior, and to asses whether it could et al., 2016) and concluded that camelina coproducts totally or partially substitute canola meal in beef can replace conventional supplements in the diets. feedlot diets. However, others described a significant decrease in dry matter intake (DMI) when a forage diet was MATERIALS AND METHODS supplemented with a limited amount of concentrate containing camelina meal (Cappellozza et al., 2012) Two experiments were carried out from October or a tendency to a decreased DMI when camelina 2017 to February 2018. In the first, two free-choice meal was used in a dairy cow diet (Hurtaud and tests were performed to ascertain if animals showed Peyraud, 2007). When comparing in situ the two a preference when they were offered two different camelina coproducts, Salas et al. (2019) concluded diets, one based on canola meal and another on that they differ in crude protein (CP) coefficient of camelina expeller. In the second, we studied the degradability, being higher in camelina expeller than effects of replacing canola meal with camelina ex- in camelina meal. This difference was confirmed peller on intake, digestibility, and feeding behavior when they were included in isoenergetic and isoni- of beef heifers fed high-concentrate diets. Animal trogenous diets based on these protein ingredients, procedures were approved by the Institutional resulting in a different dietary nitrogen flow when Animal Care and Use Committee (reference tested in an in vitro trial using a dual flow continuous CEEAH 1676) of the Universitat Autònoma de culture system, and with 8 d of incubation (Salas Barcelona (Spain) in accordance with the European et al., 2019). However, neither camelina expeller nor directive 2010/63/EU. camelina meal differed from canola meal, which was also included in this in vitro study, shows a similar Free-Choice Tests efficiency of microbial protein synthesis. This simi- larity would suggest that camelina coproducts could Twenty-four Simmental heifers [273 ±9.6 d substitute canola meal in beef cattle diets. old and with an average initial body weight (BW) Translate basic science to industry innovation 924 Salas et al. of 258.6 ±3.6 kg] were allotted in four pens, with Table 1. Ingredients and chemical composition of six heifers per pen. Pen was considered the experi- treatment diets tested in the free-choice tests1 mental unit in which the two treatments tested were CM2 CE separately offered at the same time (four replicates Ingredients, % (DM basis) per treatment). Body weight in each pen was on Corn grain 37.5 26.9 average 259.5, 256.1, 258.3, and 261.3 kg. Each pen Barley grain 25.8 36.6 of 25 m2 was equipped with two independent feed- Canola meal 15.8 — bunks located 2.5 m apart and a water trough be- Camelina expeller — 14.6 side each one. To record feed intake, each feedbunk Soybean hulls 7.7 8.9 was mounted on a waterproof digital platform scale Barley straw 10.3 10.5 (model DI-160; DIGI I’s Ltd, Maesawa-cho, Isawa- Salt 0.5 0.5 gun, Iwake, Japan). Each scale was programmed Bicarbonate 1.0 1.0 to transmit the feed weight at 5-s intervals to a Calcium carbonate 0.9 0.9 Vitamin–mineral premix3 0.4 0.4 computer with appropriate data capture software Chemical composition, % (DM basis)4 (LabView; National Instruments Corporation, DM 90.0 ± 0.365 90.4 ± 0.34 Austin, TX). Water consumption was recorded Ash 4.2 ± 0.18 4.5 ± 0.20 with direct reading flow meters (B98.32.50, Invesy CP 13.3 ± 0.72 13.2 ± 0.79 model 510 C, Tashia S.L., Artesa de Segre, Spain). EE 2.4 ± 0.05 3.4 ± 0.03 Two isoenergetic (2.8 Mcal ME/kg DM) and NDF 25.0 ± 0.64 25.3 ± 0.82 isonitrogenous (13% CP, on DM basis) concen- ADF 13.1 ± 0.48 13.6 ± 0.54 trates were formulated to meet recommendations Lignin 1.5 ± 0.09 1.7 ± 0.06 of the Fundación Española para el Desarrollo de la NFC6 55.1 ± 0.29 53.6 ± 0.42 Nutrición Animal (FEDNA, 2008) for beef cattle 1 Five percent of molasses (as-fed basis) was added to each concen- to obtain a weight gain of 1.2 kg/d. One concen- trate using a mixer feeder wagon in the second free-choice test. trate was based on canola meal (CM) as main pro- 2 CM = Total mixed ration with canola meal as main protein source; tein source and another on camelina expeller (CE). CE = Total mixed ration with camelina expeller as main protein source. The main ingredients of the concentrate, except 3 Vitamin–mineral premix (Pinsos Nutribó S.A., Sant Joan de les Abadeses, Girona, Spain) contained per kg (as fed): 8,400 IU vitamin minerals and premix, were ground through a 5-mm A, 1,680 IU vitamin D3, 39.0 mg Fe, 0.7 mg I, 0.7 mg Co, 3 mg Cu, screen. Each concentrate was manually mixed with 30 mg Mn, 85 mg Zn, 0.2 mg Se, 78 mg calcium propionate, 42 mg a mechanically chopped barley straw to achieve a malic acid, 187 mg sepiolite, 2 mg of natural extract from Castanea sativa and Vitis vinifera, 0.1 mg natural extract from Satureja hortensis, 90 to 10 concentrate to straw ratio, and diets were 9 × 109 cfu of Saccharomyces cerevisae. offered ad libitum as total mixed ration (Table 1). 4 DM = dry matter; CP = crude protein; EE = ether extract; After the first free-choice test where CM at 15.8% NDF = neutral detergent fiber; ADF = acid detergent fiber; was replaced with CE at 14.6% of inclusion [on dry NFC = nonfibrous carbohydrates. matter (DM) basis], a second one was carried out, 5 Mean ± SD. this time including in both treatment diets 5% of 6 NFC is calculated as follows: 100 − (CP + ash + NDF + EE). molasses (as-fed basis). Each free-choice test con- sisted of 9 d in which both diets were simultan- feeder were collected daily and dried in a forced air eously available. The first 3 d were for adaptation oven at 60 °C for 48 h, ground in a hammer mill to diets and feedbunks, followed by two consecu- through a 1-mm screen (P. PRAT SA, Sabadell, tive 3-d periods for pen intake control. We swapped Spain) and stored for later chemical analysis. Dry over the diet in the two feedbunks every 3 d (includ- matter content of offered feed was determined ing a position change after the adaptation period), by drying samples for 24 h in a forced air oven aiming to avoid a possible feeder location effect on at 103 °C according to AOAC (1990; ID 950.05). animal choice. After the first free-choice test, the Nitrogen content of offered feed was determined by second one was performed with the inclusion of the Kjeldahl procedure (AOAC, 1990; ID 976.05), molasses. Prior to starting the study, heifers were and ether extract (EE) was performed according fed a commercial diet where the protein sources to AOAC (1990; ID 920.30). The neutral deter- tested were not included. gent fiber (NDF) and acid detergent fiber (ADF) Feed bunks were cleaned and refusals collected contents of offered diets were determined sequen- at 0900 h each morning, and feed was offered twice tially by using an Ankom Fiber Analyzer (Ankom daily at 0930 and 1630 h, increasing the offer by at Technology, Fairport, NY) in accordance with the least 15% of the previous day’s intake in each indi- methodology provided by the company. This is vidual feed bunk. Samples of offered diets of each based on the procedure of Van Soest et al. (1991) Translate basic science to industry innovation Camelina expeller in feedlot diets 925 using a thermostable α-amylase and sodium sulfite, seven consecutive days and composited for each and expressed on an ash-free basis. The lignin pen in each sampling week (day 22 to day 28). After content of offered feed was determined after fiber collection, samples were dried in a forced air oven procedures using 72% sulfuric acid. The allyl iso- at 60 °C for 48 h, ground in a hammer mill through thiocyanate level was determined by a destilation– a 1-mm screen (P. PRAT SA, Sabadell, Spain) and volumetry procedure according to the European stored for later chemical analysis to analyze DM Directive 71/250/EEC. The erucic acid content was content, chemical composition, and particle size. analyzed by chromatography (Model 6890, Hewlett Chemical determinations were the same as de- Packard, Palo Alto, CA), according to American scribed previously. Particle size separation of offered Oil Chemists’ Society (AOCS) method CE 2–66 feed and refusals were performed using the three- (AOCS, 1998). screen Penn State Particle Separator (Nasco, Fort Atkinson, WI), obtaining four different fractions: Canola Meal Replacement with Camelina Expeller bigger than 19 mm (long); between 8 and 19 mm (medium); between 1.18 and 8 mm (short); and Twenty-four Simmental heifers (initial full smaller than 1.18 mm (fine). Sorting was calculated BW of 294.7 ± 3.7 kg) were allotted in eight pens, as the actual intake of each fraction size expressed in groups of three heifers per pen, considering pen as a percentage of the predicted intake, where pre- the experimental unit. Grouping was made with dicted intake of each fraction equals the product the aim to have a similar average pen BW. Each of as-fed intake and as-fed fraction in the diet. pen was randomly assigned to one of four ex- Values <100% indicate selective refusals, >100% is perimental treatments in a 4 × 4 replicated Latin preferential consumption, and equal to 100% is no square design. The experiment was performed in sorting (Leonardi and Armentano, 2003). Full BW four 28-d periods during which groups of three of heifers was recorded on two consecutive days at animals were allotted in each pen of 12.5 m2, the beginning and end of the experiment and at the equipped with a feed bunk and a water trough. end of each experimental period. Measurements were made in groups of three ani- Total tract apparent digestibility of treatment mals because we wanted to control intake and diets was estimated using TiO2 as external marker. feeding behavior in competitive conditions. In From day 6 to day 18 of each period, 3 kg of a con- summary, in each period of each Latin square we centrate premix containing 10 g of TiO2/kg was had one pen per treatment and three heifers per mixed with treatment diets and administered daily pen. The experiment lasted 112 d. The same pre- in each pen, following Russell et al. (2016). From viously described system was used to record feed day 16 to day 19 of each period, fecal samples of intake and water consumption. all animals were collected daily from the rectum at Four isoenergetic (2.8 Mcal ME/kg DM) and 0730 h before feed administration. To obtain a rep- isonitrogenous (13% CP, on DM basis) concentrates resentative sample of the pen, 50 g of sample from were formulated to meet Fundación Española para each animal in the same pen was taken and then el Desarrollo de la Nutrición Animal (FEDNA, mixed. Fecal samples were dried in a forced air oven 2008) recommendations for beef cattle to obtain at 60 °C for 48 h, ground in a hammer mill through a weight gain of 1.2 kg/d. Diets were designed to a 1-mm screen (P. PRAT SA, Sabadell, Spain), and contain 1) CM as main protein source and 0% stored for later chemical analysis to analyze DM, of CE (0CE), 2) 3% of CE replacing CM (3CE), organic matter (OM), CP, and NDF content. From 3) 6% of CE replacing CM (6CE), and 4) 9% of the marker concentration of offered feed and feces, CE replacing CM (9CE). The main ingredients of DM digestibility of the diets was estimated with the the concentrate, except minerals and premix, were following equation: ground through a 5-mm screen. Each concentrate DM digestibility = (1 − [TiO2 in of fered feed ]/[ TiO2 in feces]) was manually mixed with mechanically chopped × 100 barley straw to achieve a 90 to 10 concentrate to straw ratio, and diets were offered ad libitum as total mixed ration (Table 2). The concentration of the marker in samples of Feedbunks were cleaned and refusals collected offered feed and feces was determined using the pro- at 0900 h each morning, and feed was offered twice cedure outlined by Myers et al. (2004). Duplicate daily at 0930 and 1630 h, increasing the offer by 0.5-g samples were weighed in 250-mL Kjeldahl at least 15% of the previous day’s intake. Samples digestion tubes. Three and a half g of K2SO4 and of feed offered and refusals were collected daily for 0.4 g of CuSO4 and 13 mL of concentrated H2SO4 Translate basic science to industry innovation 926 Salas et al. Table 2. Ingredients and chemical composition of treatment diets with camelina expeller (CE) instead of canola meal Diets1 0CE 3CE 6CE 9CE Ingredients, % (DM basis) Corn grain 37.5 35.5 33.6 31.6 Barley grain 25.8 27.8 29.8 31.8 Canola meal 15.8 12.9 9.9 7.0 Camelina expeller — 2.7 5.4 8.1 Soybean hulls 7.7 8.0 8.2 8.4 Barley straw 10.3 10.3 10.3 10.2 Salt 0.5 0.5 0.5 0.5 Bicarbonate 1.0 1.0 1.0 1.0 Calcium carbonate 0.9 0.9 0.9 0.9 Vitamin–mineral premix2 0.4 0.4 0.4 0.4 Chemical composition, % (DM basis)3 DM 90.0 ± 0.374 90.1 ± 0.34 90.1 ± 0.34 90.4 ± 0.44 Ash 4.2 ± 0.17 4.6 ± 0.17 4.2 ± 0.17 4.6 ± 0.15 CP 13.3 ± 0.82 13.1 ± 0.89 13.3 ± 0.78 13.0 ± 0.79 EE 2.4 ± 0.02 2.8 ± 0.04 2.8 ± 0.02 3.0 ± 0.03 NDF 25.0 ± 0.61 25.2 ± 0.69 24.4 ± 0.63 24.7 ± 0.61 ADF 13.1 ± 0.53 13.2 ± 0.69 13.0 ± 0.70 12.9 ± 0.67 Lignin 1.5 ± 0.05 1.6 ± 0.06 1.7 ± 0.06 1.4 ± 0.09 NFC5 55.1 ± 0.40 54.3 ± 0.43 55.3 ± 0.39 54.7 ± 0.41 Particle size, % Long 5.32 5.59 5.74 5.71 Medium 2.37 2.11 2.26 2.14 Short 1.61 1.56 1.56 1.66 Fine 90.70 90.73 90.45 90.48 1 0CE = diet with a 0% of CE; 3CE = diet with a 3% of CE; 6CE = diet with a 6% of CE; 9CE = diet with a 9% of CE. 2 Vitamin–mineral premix (Pinsos Nutribó S.A., Sant Joan de les Abadeses, Girona, Spain) contained per kg (as fed): 8,400 IU vitamin A, 1,680 IU vitamin D3, 39.0 mg Fe, 0.7 mg I, 0.7 mg Co, 3 mg Cu, 30 mg Mn, 85 mg Zn, 0.2 mg Se, 78 mg calcium propionate, 42 mg malic acid, 187 mg sepi- olite, 2 mg of natural extract from Castanea sativa and Vitis vinifera, 0.1 mg natural extract from Satureja hortensis, 9 × 109 cfu of Saccharomyces cerevisae. 3 DM = dry matter; CP = crude protein; EE = ether extract; NDF = neutral detergent fiber; ADF = acid detergent fiber; NFC = nonfibrous carbohydrates. 4 Mean ± SD. 5 NFC is calculated as 100 − (CP + ash + NDF + EE). were added to each tube. Samples were exposed to County, Taiwan). A digital color camera (model a temperature of 420 °C for 2 h. Ten milliliters of VIVOTEK IP7142; VIVOTEK INC., Chung-HO, 30% H2O2 was added to each tube. The total liq- Taipei County, Taiwan) was set up in front of the uid weight was brought up to 100 g using distilled feeding area of each pen at a height of 3 m per- water and filtered through Whatman No. 541 filter mitting a full view of the pen. An infrared light paper to remove any precipitate. The absorbance with photoelectric cells was set up at each end was measured at 410 nm in a calibrated spectro- of the paddock to allow video recording at night photometer with working standards, prepared by (λ = 830 nm and 500 W; Dennard 2020; Dennard, adding 0, 2, 4, 6, 8, and 10 mg of TiO2. To esti- Hants, UK). To study behavioral activities, we used mate the digestibility of nutrient components, the only 1 of the 2 Latin squares included in the exper- previous formula was multiplied by the quotient iment. The behavior was video-recorded for 24 h between the concentration of the nutrient in the on 2 non consecutive days of each sampling week feces and the concentration of the nutrient in the (day 22 to day 28). In accordance with Madruga feed offered. et al. (2017a), data processing was carried out by Animal behavior was recorded using a digital a time sampling method at intervals of 5 min. The video recording device (model VS-101P VioStor behavioral activities recorded were eating and rumi- NVR; QNAP Systems Inc., Xizhi City, Taipei nating. Data for each activity are presented as the Translate basic science to industry innovation Camelina expeller in feedlot diets 927 total time, expressed in minutes, in which the ani- RESULTS AND DISCUSSION mal maintained this specific activity. An observa- tion was recorded as eating when the animal had its Free-Choice Tests muzzle in the feedbunk or was chewing or swallow- Free-choice experiments can be used to assess ing food with its head over it. Ruminating included the willingness of animals to ingest certain exper- the regurgitation, mastication, and swallowing of imental feeds when offering different feeds sep- the bolus. Eating and ruminating time, expressed arately at the same time, thus determining their as min/kg total DM and min/kg NDF, were cal- preference for, or the palatability of different feeds culated taking into account both time spent eating (Meier et al., 2012). When the first free-choice test and ruminating and total DM and NDF intake was performed, heifers showed a higher prefer- recorded. Time spent masticating resulted from the ence for CM than for CE (38.6 vs. 8.7; P < 0.001; sum of eating and ruminating activities. Table 3), and this preference was maintained throughout the day (Fig. 1). We considered three Statistical Analyses possible explanations for this result. First, it could have been a case of feed neophobia, in which ani- To analyze the preference between the diets mals eat only a small amount of a new feed when offered in the free-choice test, data were statistic- it is first offered (Provenza, 1995). Low intake ally analyzed using a mixed-effects linear regres- in feedlot diets is often observed when animals sion model from the MIXED procedure of SAS are initially received into feedlots (Zinn, 1988). (version 9.3; SAS Institute Inc., Cary, NC). The However, considering that heifers used in the pres- model contained the fixed effects of treatment ent experiment did not know either of the two main and feeder position, the random effect of pen, protein sources used, we discarded this explana- and day as a repeated measure. To analyze the tion. As a second possibility, we took into account preference between diets in each hour of the day, the antinutritional factors, which are very com- a paired t-test was used (SAS Inst. Inc.). Pen was mon in all Brassica species (Tripathi and Mishra, considered the experimental unit, and data were 2007), and their adverse effects on feed intake expressed as feed intake (as-feed basis) per pen. (D’Mello, 2000). Although ruminants are more For the Latin square experimental design, the data tolerant to glucosinolates than nonruminants, it for each pen, calculated as an average of the three is recommended not to exceed 10% of inclusion heifers per pen fed a given treatment diet at each of camelina coproducts in cattle diets (FEDNA, period, were considered the experimental unit in 2010). To discard this possible effect, we analyzed all the analyses. In each experimental period, the the content of allyl isothiocyanate and erucic acid daily mean value was calculated as the average ei- in both protein ingredients. The resulting amounts ther of 7 d for DM and nutrient intake, DM intake were 0.118 and <0.05 mg/g of allyl isothiocyanate by particle size, and sorting behavior, of 3 d for and <0.01 and 0.02 g/100 g of erucic acid for CM digestibility data, or of 2 d for behavioral activ- and CE, respectively. The content of allyl isothi- ities. The normality of the data was checked with ocyanate of CE, as a major metabolite of glu- the UNIVARIATE procedure of SAS (v. 9.3; SAS cosinolates, was below that of CM, both values Institute Inc.). All these data were statistically being under the range of values (between 0.3 and analyzed using a mixed-effects linear regression model from the MIXED procedure of SAS (ver- sion 9.3; SAS Institute Inc.). The model contained Table 3. Average as-fed feed intake (kg/pen/d) of the fixed effects of Latin square, treatment, and heifers fed canola meal and camelina expeller diets period, and the random effect of pen nested within in free-choice tests Latin square. In the case of behavioral activities, Diets1 the Latin square effect was not considered, and day Item CM CE SEM P-value was included as a repeated measure. Orthogonal Free-choice test 12 contrasts were used to determine the linear and Intake, kg/d 38.6 8.7 0.90 0.001 quadratic effects of increasing the proportion of Free-choice test 23 CE in the diet. To determine whether heifers sorted Intake, kg/d 39.1 9.8 0.66 0.001 against or for each particle size, sorting behavior 1 CM = TMR with canola meal as main protein source; CE = TMR was tested for a difference from 100 using a t-test. with camelina expeller as main protein source. Significance was declared at P < 0.05, and tenden- 2 Free-choice test with diets without molasses. cies are discussed at P < 0.10. 3 Free-choice test with diets with molasses. Translate basic science to industry innovation 928 Salas et al. 2.1 mg/g) obtained by Tripathi and Mishra (2007) change of the position of the feedbunk in the pen in different varieties of CM obtained in diverse on feed intake. oil extraction processes. Values of erucic acid pre- sented by CM and CE were below 1% of the fat Canola Meal Replacement With Camelina Expeller fraction considered to be the threshold of CM (EFSA, 2016). Therefore, the content of antinu- Dry matter intake and OM intake was unaf- tritional factors suggests that their use would not fected by diet, being on average 8.93 ± 0.214 and represent a nutritional problem for heifers. Finally, 8.53 ± 0.208 kg/d, respectively (P > 0.10, Table 4). we considered the problem of palatability, and for Because diets were formulated to be isonitroge- this reason, we decided to include in a second free- nous and with similar NDF content, intake of choice test 5% of molasses to cover up a possible CP and NDF (on average 1.17 ± 0.040 and 2.14 ± bad taste and/or bad odor. However, heifers con- 0.054 kg/d, respectively; P > 0.10, Table 4) were tinued to express a greater preference for CM than not different, in accordance with the similar DM for CE (39.1 vs. 9.8; P < 0.001; Table 3), which and OM intake. Water consumption did not differ once again was maintained throughout the day among diets (P > 0.10, Table 4). The fact that the (Fig. 2). Nevertheless, it is not possible to entirely maximum level of CE inclusion in the concentrate rule out a problem of palatability because 5% of was 9%, unlike the free-choice test where CE was molasses may have been insufficient to make CE included at 14.6%, and heifers had 3 wk of diet diet more palatable for heifers. In both free-choice adaptation, could explain the absence of palat- tests, we did not find a significant effect (P > 0.05) ability problems. These results contrast with the of either the position of the pen in the barn or the finding by Cappellozza et al. (2012), working with Figure 1. Pen feed intake over the day in free-choice test 1 without molasses added to diets with camelina expeller (CE) or canola meal (CM). Intake differed between diets: ***P < 0.001; **P < 0.01; *P < 0.05; +P < 0.10. Figure 2. Pen feed intake over the day in free-choice test 2 with molasses added to diets with camelina expeller (CE) or canola meal (CM). Intake differed between diets: ***P < 0.001; **P < 0.01; *P < 0.05; +P < 0.10. Translate basic science to industry innovation Camelina expeller in feedlot diets 929 Table 4. Intake and water consumption of heifers fed diets with different proportion of canola meal and camelina expeller as main protein source Diets1 P-value Item2 0CE 3CE 6CE 9CE SEM Linear Quadratic Intake, kg/d DM 8.88 8.90 9.06 8.89 0.214 0.780 0.553 OM 8.47 8.51 8.66 8.49 0.208 0.747 0.488 CP 1.17 1.18 1.16 1.17 0.040 0.819 0.866 NDF 2.10 2.13 2.17 2.14 0.054 0.437 0.424 Water consumption, L/day 26.95 25.40 26.81 24.92 1.120 0.161 0.918 1 0CE = diet with a 0% of CE; 3CE = diet with a 3% of CE; 6CE = diet with a 6% of CE; 9CE = diet with a 9% of CE. 2 DM = dry matter; OM = organic matter; CP = crude protein; NDF = neutral detergent fiber. Table 5. Total tract apparent digestibility of the diets with different proportions of canola meal and camelina expeller Diets1 P-value Item 2 0CE 3CE 6CE 9CE SEM Linear Quadratic Apparent digestibility, % DM DM 64.76 63.14 62.36 65.33 1.548 0.851 0.055 OM 68.59 67.25 66.97 68.85 1.092 0.883 0.055 CP 54.49 53.99 52.23 53.39 1.860 0.396 0.535 NDF 36.92 32.70 33.61 35.57 2.602 0.702 0.115 1 0CE = diet with a 0% of CE; 3CE = diet with a 3% of CE; 6CE = diet with a 6% of CE; 9CE = diet with a 9% of CE. 2 DM = dry matter; OM = organic matter; CP = crude protein; NDF = neutral detergent fiber. beef steers fed a diet based on mixed alfalfa-grass a tendency for a quadratic effect was detected in hay offered ad libitum and receiving grain-based DM and OM digestibility (P = 0.055; Table 5). This supplements, that total DMI was reduced when unclear quadratic tendency and the nonlinear effect camelina meal was included in the supplement on nutrient digestibilities would confirm those that offered (2.04 kg of DM/steer) instead of soy- were obtained in vitro in a previous experiment bean meal (2.20 kg of DM/steer). These authors (Salas et al., 2019) when canola meal and camelina included camelina meal at 6.3% (on a DM basis) expeller were tested individually as ingredients or in one of the experiments carried out and at 8.5% when these were included in a diet for beef in pro- in another. In the present study, CE was included portions similar to those used in the free-choice at 2.7% (3CE), 5.4% (6CE), and 8.1% (9CE) in the tests and using a dual flow continuous culture sys- diet, and DMI was not affected. The main differ- tem. In an in vitro experiment, also using ferment- ences between both experiments were that we used ers, Brandao et al. (2018) compared the digestibility CE instead of camelina meal and that we formu- of three diets with 0, 50, and 100% of a solvent-ex- lated a diet with a 90 to 10 concentrate to straw tracted camelina meal replacing canola meal. They ratio, while in their experiment, the amount of did not detect any effect of diet on DM, OM, concentrate was limited, and animals consumed CP, and ADF digestibility, but observed a linear forage ad libitum. In fact, Cappellozza et al. decrease in NDF digestibility. These authors sug- (2012) reported that camelina meal impaired both gested that increased inclusion of camelina meal forage intake and total DMI. In agreement with mainly affected hemicellulose digestibility, which our results, Lawrence et al. (2016) did not report they related to the decreased acetate proportion differences in DMI when growing heifers were fed in diets containing camelina meal detected in their diets containing 60% grass hay and 40% concen- experiment. This different result could be related trate mix, in which either 10% camelina meal, 10% to the type of diet used. Although Brandao et al. distillers dried grains with solubles, or 10% linseed (2018) formulated diets for dairy cows that con- meal was used as an ingredient in the concentrate. tained 55% hay and 45% concentrate, we designed Total tract apparent digestibility of DM, OM, diets for fattening beef with 10% barley straw and CP, and NDF was linearly unaffected by diet, but 90% concentrate. Translate basic science to industry innovation 930 Salas et al. When different particle sizes of diets were sep- remaining particle size did not differ among diets. arated using the three-screen Penn State Particle This result corresponds with the tendency for a lin- Separator, the proportions of each particle size ear increase of sorting extension for this particle size were similar among diets, being on average 5.6 ± (P = 0.07) and with a sorting behavior against long 0.08, 2.2 ± 0.05, 1.6 ± 0.02, and 90.6 ± 0.06 % for particle size (Table 7; P < 0.05) detected in 0CE and long, medium, short, and fine particles, respec- 3CE, but not in 6CE and 9CE, where no sorting was tively (Table 2). However, intake of long particle observed (Table 7). In addition, sorting for fine par- size increased linearly as CE proportion increased ticle size was detected in heifers fed all diets except in the diet (Table 6; P < 0.015), whereas intake of for 6CE (Table 7), this particle size corresponding Table 6. Intake by particle size of heifers fed diets with different proportions of canola meal and camelina expeller Diets1 P-value Item 0CE 3CE 6CE 9CE SEM Linear Quadratic Intake by particle size2, kg/d Long 0.35 0.44 0.48 0.46 0.043 0.015 0.063 Medium 0.09 0.10 0.11 0.11 0.026 0.674 0.835 Short 0.08 0.08 0.10 0.09 0.017 0.310 0.768 Fine 8.25 8.28 8.38 8.24 0.203 0.901 0.549 1 0CE = diet with a 0% of CE; 3CE = diet with a 3% of CE; 6CE = diet with a 6% of CE; 9CE = diet with a 9% of CE. 2 Particle size determined by Penn State Particle Separator. Table 7. Effect of diet on sorting behavior of heifers fed diets with different proportions of canola meal and camelina expeller Diets1 P-value Item 0CE 3CE 6CE 9CE SEM Linear Quadratic Particle size2 Long 78.26* 3 87.75* 91.18 90.90 7.138 0.070 0.316 Medium 47.85 54.82 58.65 56.48 12.808 0.450 0.600 Short 56.67 60.36 65.71 62.70 11.304 0.499 0.662 Fine 103.21* 102.56* 102.18 102.46* 0.840 0.295 0.412 1 0CE = diet with a 0% of CE; 3CE = diet with a 3% of CE; 6CE = diet with a 6% of CE; 9CE = diet with a 9% of CE. 2 Particle size determined by Penn State Particle Separator. 3 Values equal to 100% indicate no sorting, <100% selective refusals (sorting against), and >100% preferential consumption (sorting for). *Statistical differences from 100% are expressed as follows: P < 0.05. Table 8. Effect of diet on animal behavior of heifers fed diets with different proportions of canola meal and camelina expeller Diets1 P-value Item 0CE 3CE 6CE 9CE S.E.M. Linear Quadratic Eating Min/d 95.5 91.8 100.2 91.9 7.27 0.907 0.590 Min/kg DM 10.4 10.0 10.8 10.3 0.55 0.809 0.993 Min/kg NDF 43.6 41.5 45.0 44.1 2.35 0.511 0.715 Ruminating Min/d 361.1 369.3 390.2 376.2 21.50 0.381 0.509 Min/kg DM 40.1 40.0 42.0 42.9 2.79 0.254 0.802 Min/kg NDF 167.2 165.6 175.9 183.7 11.91 0.131 0.585 Chewing Min/d 456.6 461.1 490.4 468.1 21.69 0.421 0.452 Min/kg DM 50.5 50.0 52.7 53.2 2.94 0.260 0.813 Min/kg NDF 210.8 207.0 220.9 227.8 12.55 0.121 0.558 1 0CE = diet with a 0% of CE; 3CE = diet with a 3% of CE; 6CE = diet with a 6% of CE; 9CE = diet with a 9% of CE. Translate basic science to industry innovation Camelina expeller in feedlot diets 931 essentially with the concentrate ingredient of diets. Regional Development Fund (Research Project Sorting behavior has been reported in growing RTC-2015-265-5) is acknowledged. calves (Miller-Cushon et al., 2013; Groen et al., 2015; Gordon and DeVries, 2016) and growing CONFLICT OF INTEREST heifers (Greter et al., 2008; DeVries et al., 2014). The authors declare no conflict of interest. Specifically, sorting behavior with preferential con- sumption for concentrate in total mixed ration LITERATURE CITED (TMR) in beef heifers fed high-concentrate diets was described by Madruga et al. (2017b), confirm- AOAC. 1990. Official methods of analysis. 15th ed. Assoc. Off. ing the result obtained in the present experiment. Anal. Chem., Arlington, VA. AOCS. 1998. Official methods and recommended practices of Both the greater intake of long particles and the AOCS. American Oil Chemists’ Society, Champaign, the lack of sorting for this particle size recorded in IL. diets with a greater proportion of CE would indi- Brandao, V. L. N., L. G. Silva, E. M. Paula, H. F. Monteiro, cate that heifers fed these diets needed to consume X. Dai, A. L. J. Lelis, A. Faccenda, S. R. Poulson, and more barley straw. This might suggest that, in ac- A. P. Faciola. 2018. Effects of replacing canola meal with solvent-extracted camelina meal on microbial fermenta- cordance with the free-choice tests, animals de- tion in a dual-flow continuous culture system. J. Dairy Sci. tected some palatable characteristics that induced 101:9028–9040. doi:10.3168/jds.2018-14826 them to intake more straw than animals fed 0CE. Cappellozza, B. I., R. F. Cooke, D. W. Bohnert, G. Cherian, Considering these results, differences in the chewing and J. A. Carroll. 2012. Effects of camelina meal supple- activity of heifers fed these diets could be expected. mentation on ruminal forage degradability, performance, However, it was observed that the small increase and physiological responses of beef cattle. J. Anim. Sci. 90:4042–4054. doi:10.2527/jas.2011-4664 in the intake of longer particles was not enough to Colombini, S., G. A. Broderick, I. Galasso, T. Martinelli, promote differences in chewing activity. Time spent L. Rapetti, R. Russo, and R. Reggiani. 2012. Evaluation eating and ruminating, expressed as min/d, min/kg of Camelina sativa (L.) Crantz meal as an alternative DM, and min/kg NDF, was unaffected by diet (P > protein source in ruminant rations. J. Sci. Food Agric. 0.10; Table 8), and only a numerical linear increase 94:736–743. doi:10.1002/jsfa.6408 DeVries, T. J., T. Schwaiger, K. A. Beauchemin, and in time spent ruminating and chewing was observed G. B. Penner. 2014. The duration of time that beef cat- as the CE proportion increased, when expressed as tle are fed a high-grain diet affects feed sorting behavior min/kg NDF with a P-value of 0.131 and 0.121, both before and after acute ruminal acidosis. J. Anim. 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Fundación Española para el Desarrollo de and digestibility of these high-concentrate diets la Nutrición Animal, Madrid, Spain, 502 pp. www.funda- offered to beef heifers were unaffected. However, cionfedna.org. (In Spanish) intake of long particle size increased as camelina FEDNA. 2008. Necesidades nutricionales para rumiantes de cebo: expeller proportion increased, and heifers fed at normas FEDNA. Fundación Española para el Desarrollo 0% and 3% of camelina expeller inclusion sorted de la Nutrición Animal. http://www.fundacionfedna.org/ against long particle size. The fact that this size cor- sites/default/files/NORMAS_RUMIANTES_2008.pdf. (Accessed January 2019). (In Spanish) responds mainly with barley straw, we may surmise Gordon, L. J., and T. J. DeVries. 2016. 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