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Animal Science Journal (2014) 85, 751–756
doi: 10.1111/asj.12196
ORIGINAL ARTICLE Determination of the net energy content of canola meal from Brassica napus yellow and Brassica juncea yellow fed to growing pigs using indirect calorimetry* Jung Min HEO,1,2 Deborah ADEWOLE1 and Martin NYACHOTI1 1
Department of Animal Science, University of Manitoba, Winnipeg, Manitoba, Canada; and 2Department of Animal Science and Biotechnology, Chungnam National University, Daejeon, South Korea
ABSTRACT The net energy (NE) content of canola meals (CM; i.e. Brassica napus yellow and Brassica juncea yellow) in growing pigs was determined using an indirect calorimetry chamber or published prediction equations. The study was conducted as a completely randomized design (n = 6), with (i) a basal diet and (ii) 2 diets containing 700 g/kg of the basal diet and 300 g/kg of either of the two varieties of CM. A total of 18 growing barrows were housed in metabolism crates for the determination of digestible (DE) and metabolizable (ME) energy. Thereafter, pigs were transferred to the indirect calorimetry chamber to determine heat production (HP). The NE contents of diets containing Brassica napus yellow and Brassica juncea yellow determined with the direct determination technique and prediction equations were 9.8 versus 10.3 MJ/kg dry matter (DM) and 10.2 versus 10.4 MJ/kg DM, respectively. Retained energy (RE) and fasting heat production (FHP) of diets containing Brassica napus yellow and Brassica juncea yellow were 5.5 versus 5.7 MJ/kg and 4.3 versus 4.5 MJ/kg, respectively, when measured with the direct determination technique and prediction equations. The NE contents of Brassica napus yellow and Brassica juncea yellow were determined to be 8.8 and 9.8 MJ/kg DM, respectively, using the direct determination technique.
Key words: canola meal, indirect calorimetry, net energy, pig.
INTRODUCTION Due to an increase in domestic canola crushing capacity, canola meal (CM) production in Canada is expected to expand. Although CM has considerable crude protein content and a well-balanced amino acide profile, its high fiber content limits its use in pig diets because of limited energy availability (Bell 1993). Furthermore, the energy contents of fiber- and protein-rich feeds are often overestimated when using the digestible energy (DE) and metabolizable energy (ME) systems (Noblet et al. 1994a). As a consequence, canola breeders have developed new varieties with high energy content and lower fiber contents (Simbaya et al. 1995; Slominski 1997). Recent studies have shown that CM (i.e. solvent-extracted and expeller-pressed CM) can be used in weaned pig diets up to 200 g/kg without compromising growth performance when diets were formulated to equivalent net energy (NE) content and standard ileal digestible lysine/MJ NE (Landero et al. 2011, 2012). These results support the use of the NE system in formulating © 2014 Japanese Society of Animal Science
swine diets, especially for those utilizing high fiber ingredients. Despite the demonstrated superiority of the NE system over the DE and ME systems, its utility in swine diet formulation has not gained widespread application in some regions, including North America, Asia and Australasia. Although published prediction equations (Noblet et al. 1994a,b) are used to estimate the NE content in swine diets and this is supported by results of some recent studies (Ayoade et al. 2012), others have shown that predicted values may not Correspondence: Martin Nyachoti, Department of Animal Science, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada. (Email: [email protected]) *An abstract of this work has been published in Journal of Animal Science Supplements 2. pp 88–89, 2012 as: Use of indirect calorimetry to determine the net energy (NE) content of canola meals in growing pigs. J. M. Heo, D. Adewole and C. M. Nyachoti. Received 30 September 2013; accepted for publication 7 January 2014.
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always agree with empirical data (Kil et al. 2011). Also, prediction equations are optimized to evaluate NE content in complete diets and therefore it might be challenging to acquire the NE values from the prediction equations for individual ingredients (NRC 2012). Thus, there is a need to further evaluate the NE values of swine feeds and feed ingredients using prediction equations and empirical measurements so as to guide its utility in formulating swine diets. Furthermore, the NE content of the new varieties of yellow seeded canola (i.e. Brassica napus yellow and Brassica juncea yellow) developed to yield CM with superior nutritive value for swine has not been extensively evaluated. In the present study we tested the hypotheses that the NE contents of CM derived from Brassica napus yellow and Brassica juncea yellow were similar and that values obtained with the direct determination technique and published prediction equations would differ. Thus, the objective of the present study was to determine the NE content of CM derived from Brassica napus yellow and Brassica juncea yellow using the direct determination technique (i.e. indirect calorimetry) in growing pigs, and to compare the NE values obtained with the direct determination technique and published prediction equations.
MATERIALS AND METHODS The experimental protocol used in the present study was reviewed and approved by the Animal Care Committee of the University of Manitoba. Animals were cared for according to the guidelines of the Canadian Council on Animal Care (CCAC 2009).
Animals A total of 18 growing barrows (Large White × Landrace × Duroc) with an average initial body weight (BW) of 14.9 ± 0.11 kg (mean ± SEM) were acquired from the Gleanlea Swine Research Unit, University of Manitoba. Pigs were individually housed for 15 days in adjustable metabolism crates (1.80 × 0.6 m) with smooth transparent plastic sides and plastic-covered expanded metal sheet flooring in a temperature-controlled room (22 ± 2°C).
Diets The basal diet was formulated based on corn-soybean meal to meet NRC (1998) requirements for growing pigs. Two test diets were formulated with 700 g/kg of the basal diet and 300 g/kg of CM (i.e. Brassica napus yellow or Brassica juncea yellow; Table 1).
Experimental design and procedures This study was conducted in two consecutive periods (10 pigs for period 1 and 8 pigs for period 2) using the same facility and similar experimental conditions and procedures because only two indirect calorimetry chambers were available at the time for the current study. Pigs were assigned to one of three experimental diets in a completely randomized design to give six replicates per diet. In each period, pigs were housed in metabolism crates and fed experimental diets for 15 days, © 2014 Japanese Society of Animal Science
Table 1 Composition and analysis of experimental diet (as-fed basis)
Item
Diets
Ingredients, g/kg Corn Soybean meal Brassica napus yellow Brassica juncea yellow Vegetable oil Salt Monocalcium phosphate Limestone Vitamin/mineral premix† Lysine-HCl DL-methionine L-threonine Calculated composition Crude protein, g/kg DE, MJ/kg ME, MJ/kg NE, MJ/kg SID lysine, g/MJ DE NDF, g/kg Crude fat, g/kg Analysed composition Crude protein, g/kg Dry matter, g/kg GE, MJ/kg Ash, g/kg NDF, g/kg ADF, g/kg Ether extract, g/kg Starch, g/kg
Basal
BNY
BJY
679.1 272.5
475.2 190.8 300.0
475.2 190.8
12.5 5.0 9.6 10.5 10.0 0.60 0.10 0.10
8.8 3.5 6.7 7.4 7.0 0.40 0.10 0.10
300.0 8.8 3.5 6.7 7.4 7.0 0.40 0.10 0.10
177 14.5 13.8 10.0 0.60 101 31
230 13.8 13.2 9.7 0.74 147 27
242 14.3 13.6 9.9 0.73 119 29
174 883 16.3 51 118 18 39 386
231 890 16.6 54 199 23 34 280
248 894 16.9 56 168 28 32 290
†Provided the following nutrients (per kg of air-dry diet): vitamins: A, 2000 IU, D3 200 IU, E, 40 mg, K, 2 mg, B1, 1.5 mg, B2, 7 mg, B6, 2.5 mg, B12, 25 μg, calcium pantothenate, 14 mg, folic acid, 1 mg, niacin, 21 mg, biotin, 70 μg. Minerals: Cu, 10 mg (as copper sulphate), iodine, 0.4 mg (as potassium iodine), iron, 120 mg (as ferrous sulphate), Mn, 10 mg (as manganous oxide), Se, 0.3 mg (as sodium selenite), Zn, 110 mg (as zinc oxide). BNY, Brassica napus yellow; BJY, Brassica juncea yellow; ADF, acid detergent fibre; DE, digestible energy; GE, gross energy; ME, metabolisable energy; NDF, neutral detergent fibre; NE, net energy; SID, standard ileal digestible.
including 10 days for adaptation to feed and environmental conditions and 5 days for total collection of feces and urine to measure DE and ME contents. Pigs were fed their respective diets at 2.3 MJ of ME/kg BW0.6 per day based on BW on days 1, 5 and 10 which was close to ad libitum intake (after Noblet et al. 1994b). During the study, pigs were fed at 08.30 hours and trained to consume their daily feed allowance within 1 h. Pigs had unlimited access to water via a low-pressure nipple throughout the study. During the last 5 days of each feeding period, total fecal and urine collection for the estimation of DE and ME was performed as previously described (Woyengo et al. 2010). On day 16, two pigs were transferred to the indirect calorimetry chambers (1.22 × 0.61 × 0.91 m; Columbus Instruments, Columbus, OH, USA) for 36 h of heat production (HP) and fasting HP (FHP) measurement based on O2 consumption, CO2 production and urine output. The following sets of two pigs were Animal Science Journal (2014) 85, 751–756
NET ENERGY CONTENT OF CANOLA MEAL
transferred to the indirect calorimetry chambers every 2 days (i.e. days 18, 20, 22, 24 and 26). Pigs were brought into the indirect calorimetry chambers within 1 h after consumption of their daily ration and HP was measured continuously for 24 h followed by 12 h of FHP measurement. Pigs were allotted to one of two indirect calroimetry chambers in a completely randomized design to avoid possible confounding effects. Fresh water was available in the indirect calorimetry chambers at all times and urine voided during the 24 h and 12 h period were collected separately, weighed, sub-sampled and stored at −20°C until required for N analysis. Temperature within the indirect calorimetry chambers was maintained at 22 ± 1°C and personnel movement around the indirect calorimetry chambers was limited to avoid disturbing pigs during HP and FHP measurements.
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The NE contents of the basal and CM containing diets are calculated according to the equations established by Noblet et al. (1994b):
NE (MJ /kg DM) = 0.843 × DE (MJ /kg DM) − 463
(1)
NE (MJ/kg DM) = 0.703 × DE (MJ /kg DM) + 1.58 × ether extract (% DM) + 0.47 × starch (% DM) (2) − 0.97 × crude protein (% DM) − 0.98 × crude fiber (%DM) NE (MJ /kg DM) = 0.700 × DE (MJ /kg DM) + 1.61 × ether extract (% DM) + 0..48 × starch (% DM) − 0.91 × crude (3) protein (% DM) − 0.87 × acid deterrgent fiber (% DM) The NE of test CM was calculated as follows:
Sample preparation and chemical analyses Dry matter (DM) was determined according to AOAC (1990; method 925.09) and gross energy (GE) content was measured using an adiabatic bomb calorimeter (model 6300; Parr Instrument, Moline, IL, USA), which had been calibrated using benzoic acid as a standard. N content was determined using the combustion method (990.03; AOAC 1990) using the LECO (model CNS-2000; LECO Corp., St. Joseph, MI, USA) N analyzer. Acid detergent fiber (ADF) and neutral detergent fiber (NDF) contents were determined according to the method of Goering and Van Soest (1970) and ash content was determined according to AOAC (1990; method 942.05). Ether extract (EE) was determined according to AOAC (1990; method 920.39). Starch content was measured according to the AOAC (1990; method 996.11) using an assay kit (Megazyme Total Starch assay kit; Megazyme International Ltd, Wicklow, Ireland). Non-starch polysaccharides were analyzed using gas-liquid chromatography (component neutral sugars) using an SP-2340 column and a Varian CP3380 gas chromatograph (Varian Inc., Palo Alto, CA, USA) and by colorimetry (uronic acids) using a Biochrom Ultrospec 50 (Biochrom Ltd, Cambridge, England) and the procedure described by Englyst and Cummings (1988) with some modifications (Slominski & Campbell 1990). The urine samples collected from the metabolism crates were analyzed for DM and GE. For the DM of urine, 1 mL of each sample was mixed with 0.5 g of cellulose and the weight of the resulting mixture recorded. The urine-cellulose mixtures together with samples of pure cellulose were dried in an oven at 50°C for 24 h. The GE was then determined on the dried urine-cellulose mixtures as described above and samples of pure cellulose, and the contents of the same in urine were calculated by the difference method (Fleischer et al. 1981).
Calculations Heat production, FHP, retained energy (RE) and NE were calculated using the following equations:
HP (MJ/kg DM) = [3.87 × O2(L) + 1.20 × CO2(L) − 1.43 × urinary N (g)]/dry matter intake (DMI)
(Brouwer 1965) RE (MJ /day) = [ME (MJ/day) − HP (MJ /day)]/DMI NE (MJ /kg DM) = (RE MJ /day + FHP MJ /day)/DMI Animal Science Journal (2014) 85, 751–756
NETest canola meal(MJ/kg DM) = Basal diet NE − Basal diet NE − Test diet NE/0.3
Statistical methods Treatment effects were evaluated univariately in a normal mixed-liner model using the GLM procedures of SPSS (version 18.0, SPSS Inc., Chicago, IL, USA). The pig was considered as the experimental unit for all measurements and the model included dietary treatments. The experimental period was used as a random factor. Statistical significance was accepted at P < 0.05.
RESULTS All animals adapted well to their respective diets and environmental conditions, remained healthy and readily consumed their daily feed allowance during the experimental period. Nitrogen balance and HP and FHP values obtained with the direct determination technique are presented in Table 2. Nitrogen retention for pigs fed Brassica napus yellow- and Brassica juncea yellow-containing diets were 19.4 and 22.0 g/day, respectively. Corresponding values for HP were 7.3 and 7.8 MJ/kg, respectively. The energy balance of pigs and energy values of experimental diets and CM determined with the direct determination technique and prediction equations are presented in Table 3. Using the direct determination technique, the DE, ME and NE contents in Brassica napus yellow and Brassica juncea yellow were 13.6, 12.6 and 8.8 and 14.2, 13.5 and 9.8 MJ/kg DM, respectively. Respective values obtained with published prediction equations were 13.8, 12.8 and 8.1 and 15.0, 13.9 and 9.4 MJ/kg DM for Brassica napus yellow and Brassica juncea yellow, respectively. The RE and FHP of Brassica napus yellow- and Brassica juncea yellow-containing diets were 5.5 versus 5.7 MJ/kg and 4.3 versus 4.5 MJ/kg, respectively. The NE contents of diets containing Brassica napus yellow and Brassica juncea yellow were 9.8 and 10.2 MJ/kg DM and the NE contents of Brassica napus yellow and © 2014 Japanese Society of Animal Science
754 J. M. HEO et al.
Table 2 Nitrogen balance in pigs fed diets containing canola meals determined with direct determination technique
Item
Nitrogen balance Nitrogen ingested, g/day Nitrogen output in faces, g/day Nitrogen digested, % Nitrogen retained, g/day Urine nitrogen excretion, g/day
Diets Basal
Brassica napus yellow
Brassica juncea yellow
18.3 4.3 76.6 14.0 4.8
27.0 7.6 71.8 19.4 4.3
29.1 7.2 75.3 22.0 4.7
SEM
P-value
0.33 0.19 0.70 0.27 0.42
0.001 0.001 0.036 0.001 0.878
Table 3 Energy balance of pigs and energy values of experimental diets and canola meals determined with direct determination technique using the indirect calorimeter chamber and prediction equations using chemical characteristics
Item
Direct determination technique DE of diets, MJ/kg DM DE of canola meal, MJ/kg DM ME of diets, MJ/kg DM ME of canola meal, MJ/kg DM Heat production, MJ/kg Fasting heat production, MJ/kg Retained energy, MJ/kg NE of diets, MJ/kg DM NE of canola meal, MJ/kg DM Prediction equations provisions DE of diets, MJ/kg DM† DE of canola meal, MJ/kg DM† ME of diets, MJ/kg DM2 ME of canola meal, MJ/kg DM‡ NE of diets, MJ/kg DM§ NE of canola meal, MJ/kg DM§
Diets
SEM
P-value
Basal
Brassica napus yellow
Brassica juncea yellow
15.5 – 15.1 – 8.4 4.5 6.1 10.6 –
14.8 13.6 14.3 12.6 7.3 4.3 5.5 9.8 8.8
14.9 14.2 14.2 13.5 7.8 4.5 5.7 10.2 9.8
0.10 0.35 0.10 0.31 0.18 0.90 0.29 0.25 0.58
0.021 0.458 0.004 0.213 0.091 0.826 0.645 0.484 0.466
15.0 – 14.6 – 11.0 –
14.7 13.8 14.2 12.8 10.3 8.1
15.1 15.0 14.5 13.9 10.4 9.4
0.79 0.89 0.04 0.06 0.62 0.23
0.980 0.546 0.029 0.001 0.007 0.052
†Mean of prediction equations by Noblet and Perez (1993), where DE = 949 + (0.789 × GE) – (43 × % Ash) – (41 × % NDF) and DE = 4151 – (122 × % Ash) + (23 × % CP) + (38 × % EE) – (64 × % CF). ‡Mean of prediction equations by May and Bell (1971) and Noblet et al. (1989) and Noblet and Perez (1993), where ME = DE × [1.012 – (0.0019 × % CP)], ME = DE × [0.998 – (0.002 × % CP)] and ME = DE × [1.003 – (0.0021 × % CP)], respectively. §Mean of prediction equations by Noblet et al. (1994a), where NE = 0.843 × DE – 463, NE = 0.703 × DE + 1.58 × (%) EE + 0.47 × (%) starch – 0.97 × (%) CP – 0.98 × (%) CF and NE = 0.700 × DE + 1.61 × (%) EE + 0.48 × (%) starch – 0.91 × (%) CP − 0.87 × (%) ADF. BW, body weight; DE, digestible energy; ME, metabolisable energy; SEM, pooled standard error of mean; GE, gross energy; NDF, neutral detergent fiber; CP, crude protein; EE, ether extract; CF, crude fiber; ADF, acid-detergent fiber.
Brassica juncea yellow were determined to be 8.8 and 9.8 MJ/kg DM, respectively, by the direct determination technique. However, the average NE contents of Brassica napus yellow and Brassica juncea yellow determined using published prediction equations were 8.1 and 9.4 MJ/kg DM, respectively.
DISCUSSION In the present study, we have carefully selected the three prediction equations based on the DE system because the DE values are extensively available and could be effortlessly found for most feed ingredients, particularly in North America, Asia and Australasia and these equations were highly correlated (r2 = 0.910.97). In this light, we have used the mean NE values obtained from these prediction equations to compare © 2014 Japanese Society of Animal Science
with values obtained from the direct determination technique. The calculated NE content of experimental diets used in the present study were closer to the values obtained using the direct determination technique than values obtained using prediction equations reported by Noblet et al. (1994b) (see Tables 1 and 3). Nevertheless, the calculated contents of DE and ME were higher compared to the values obtained from either of the two techniques used in the current study. This discrepancy could be due to the high protein and fiber contents of experimental diets which are known to overestimate energy values when expressed on a DE and ME system compared with energy values measured in feed containing high contents of starch and fat in pigs (Noblet et al. 1994a). The analyzed gross energy, ash, EE and NDF content of test ingredients (i.e. Brassica napus yellow and Brassica juncea yellow) Animal Science Journal (2014) 85, 751–756
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were similar to reported values (e.g. Slominski et al. 1999; Montoya & Leterme 2009), but the crude protein content of the Brassica napus yellow was lower compared with values (393 vs. 466 and 519 g/kg DM, respectively) reported in the above cited studies. We suspected that this response could be attributed to excessive toasting of Brassica napus yellow because it has been reported that increasing the duration of toasting had a negative impact on soluble protein content (Barac´ & Stanojevic´ 2005). Likewise, van Barneveld et al. (1994) reported that dietary inclusion of heat-treated ingredients had significant reductions in protein deposition, ileal-digestible lysine retention and growth performance in growing pigs. This could have contributed to the relatively lower NE content of Brassica napus yellow independent of the determination techniques (i.e. direct determination vs. prediction equation; 8.8 vs. 8.1 MJ/kg DM, respectively) compared with a value of 11.9 MJ/kg DM for the same canola meal type published elsewhere (Montoya & Leterme 2009). However, the NE contents of the Brassica juncea yellow determined in this study using both techniques (i.e. direct determination vs. prediction equation; 9.8 vs. 9.4 MJ/kg DM) were comparable with values of 9.4 MJ/kg DM reported by Montoya and Leterme (2009). In addition, the NE content of Brassica juncea yellow is calculated to be 9.6 MJ/kg DM when the same prediction equation was applied with the cited study (i.e. equation 3); however, it was not possible in the present study to reconcile the accuracy between published equations used as these were developed using complete diets with various inclusion levels of ingredients. In the present study, the NE content determined using the direct determination technique was approximately 5.9% higher than the value calculated using the prediction equations (Noblet et al. 1994b). It is known that energy utilization is greater in mature pigs (Noblet & Shi 1993; Le Goff & Noblet 2001), and abnormality is essential when applying this theory because pigs used in the present study were lighter (about 26 kg) than those used (about 35–45 kg) in the studies of Noblet et al. (1994a, b). However, feeding levels, digestive physiological responses and digestive utilization of diets in pigs at around 25 to 50 kg would not relatively differ (NRC 2012). Furthermore, it is likely that the NE values of diets and ingredients could differ due to genetics, methodologies used to measure RE, feeding strategies and environmental conditions in which pigs are kept (Boisen & Verstegen 1998). In addition, the discrepancy could be due to those prediction equations per se because these were derived using complete diets. The NE content in Brassica napus yellow was lower compared with Brassica juncea yellow regardless of the NE determination technique used. This could be attributed to the lower crude protein and starch content but Animal Science Journal (2014) 85, 751–756
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Table 4 Analysed chemical composition of ingredients (g/kg, as-fed)
Item
Brassica napus yellow
Brassica juncea yellow
Crude protein Dry matter GE, MJ/kg Ash NDF ADF Crude fat Ether extract Starch Non-phytate phosphorus Total NSP
355 903 17.8 73 253 64 82 21 4 4.0 217
394 901 17.8 70 159 58 69 23 22 3.9 250
ADF = acid detergent fibre; GE = gross energy; NDF = neutral detergent fibre; NSP = non-starch polysaccharides.
higher NDF content of Brassica napus yellow than in Brassica juncea yellow (Table 4). It is known that dietary fiber (e.g. non-starch polysaccharides and NDF) is less digestible (< 50%) than other nutrients (e.g. crude protein and starch: 80–100%; Noblet & van Milgen 2004). Likewise, nitrogen balance was significantly lower in Brassica napus yellow compared to pigs fed diet containing Brassica juncea yellow that also could have contributed to lower NE content in Brassica napus yellow compared with pigs fed diet containing Brassica juncea yellow (Table 2). Nevertheless, we have used different levels of vegetable oil content in the experimental diets because only basal diet included vegetable oil. Nonetheless, Kil et al. (2011) demonstrated that the NE content of a corn-soybean meal containing 5% of soybean oil and the counterpart containing 10% of soybean oil were not affected by the content of dietary lipids (2206 vs. 2318 kcal/kg, respectively).
Conclusions The NE contents of Brassica napus yellow and Brassica juncea yellow were 8.8 versus 9.8 and 8.1 versus 9.4 MJ/kg between the direct determination technique using the indirect calorimetry chamber and prediction equations using chemical characteristics, respectively. This study showed that there is discrepancy of approximately 5.9% in the NE content between the NE determination techniques, suggesting that great variation in the NE content of diets and ingredients could be ascribed to inherent factors (i.e. BW, gender and feeding level) in relation to using different methodologies. Furthermore, research is warranted to examine the effect of the NE evaluated between the direct determination technique and prediction equations on growth performance in various BW ranges of pigs. © 2014 Japanese Society of Animal Science
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ACKNOWLEDGMENTS The authors thank Mr. Robert Stuski, Mr. Darwin Ramos, Mr. Atanas Karamanov and Ms. Lisa Rigaux for technical assistance. This research was supported by Canola Council of Canada through the Canola Science Cluster.
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Animal Science Journal (2014) 85, 751–756
Animal Science Journal (2014) 85, 751–756
doi: 10.1111/asj.12196
ORIGINAL ARTICLE Determination of the net energy content of canola meal from Brassica napus yellow and Brassica juncea yellow fed to growing pigs using indirect calorimetry* Jung Min HEO,1,2 Deborah ADEWOLE1 and Martin NYACHOTI1 1
Department of Animal Science, University of Manitoba, Winnipeg, Manitoba, Canada; and 2Department of Animal Science and Biotechnology, Chungnam National University, Daejeon, South Korea
ABSTRACT The net energy (NE) content of canola meals (CM; i.e. Brassica napus yellow and Brassica juncea yellow) in growing pigs was determined using an indirect calorimetry chamber or published prediction equations. The study was conducted as a completely randomized design (n = 6), with (i) a basal diet and (ii) 2 diets containing 700 g/kg of the basal diet and 300 g/kg of either of the two varieties of CM. A total of 18 growing barrows were housed in metabolism crates for the determination of digestible (DE) and metabolizable (ME) energy. Thereafter, pigs were transferred to the indirect calorimetry chamber to determine heat production (HP). The NE contents of diets containing Brassica napus yellow and Brassica juncea yellow determined with the direct determination technique and prediction equations were 9.8 versus 10.3 MJ/kg dry matter (DM) and 10.2 versus 10.4 MJ/kg DM, respectively. Retained energy (RE) and fasting heat production (FHP) of diets containing Brassica napus yellow and Brassica juncea yellow were 5.5 versus 5.7 MJ/kg and 4.3 versus 4.5 MJ/kg, respectively, when measured with the direct determination technique and prediction equations. The NE contents of Brassica napus yellow and Brassica juncea yellow were determined to be 8.8 and 9.8 MJ/kg DM, respectively, using the direct determination technique.
Key words: canola meal, indirect calorimetry, net energy, pig.
INTRODUCTION Due to an increase in domestic canola crushing capacity, canola meal (CM) production in Canada is expected to expand. Although CM has considerable crude protein content and a well-balanced amino acide profile, its high fiber content limits its use in pig diets because of limited energy availability (Bell 1993). Furthermore, the energy contents of fiber- and protein-rich feeds are often overestimated when using the digestible energy (DE) and metabolizable energy (ME) systems (Noblet et al. 1994a). As a consequence, canola breeders have developed new varieties with high energy content and lower fiber contents (Simbaya et al. 1995; Slominski 1997). Recent studies have shown that CM (i.e. solvent-extracted and expeller-pressed CM) can be used in weaned pig diets up to 200 g/kg without compromising growth performance when diets were formulated to equivalent net energy (NE) content and standard ileal digestible lysine/MJ NE (Landero et al. 2011, 2012). These results support the use of the NE system in formulating © 2014 Japanese Society of Animal Science
swine diets, especially for those utilizing high fiber ingredients. Despite the demonstrated superiority of the NE system over the DE and ME systems, its utility in swine diet formulation has not gained widespread application in some regions, including North America, Asia and Australasia. Although published prediction equations (Noblet et al. 1994a,b) are used to estimate the NE content in swine diets and this is supported by results of some recent studies (Ayoade et al. 2012), others have shown that predicted values may not Correspondence: Martin Nyachoti, Department of Animal Science, University of Manitoba, Winnipeg, Manitoba R3T 2N2, Canada. (Email: [email protected]) *An abstract of this work has been published in Journal of Animal Science Supplements 2. pp 88–89, 2012 as: Use of indirect calorimetry to determine the net energy (NE) content of canola meals in growing pigs. J. M. Heo, D. Adewole and C. M. Nyachoti. Received 30 September 2013; accepted for publication 7 January 2014.
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always agree with empirical data (Kil et al. 2011). Also, prediction equations are optimized to evaluate NE content in complete diets and therefore it might be challenging to acquire the NE values from the prediction equations for individual ingredients (NRC 2012). Thus, there is a need to further evaluate the NE values of swine feeds and feed ingredients using prediction equations and empirical measurements so as to guide its utility in formulating swine diets. Furthermore, the NE content of the new varieties of yellow seeded canola (i.e. Brassica napus yellow and Brassica juncea yellow) developed to yield CM with superior nutritive value for swine has not been extensively evaluated. In the present study we tested the hypotheses that the NE contents of CM derived from Brassica napus yellow and Brassica juncea yellow were similar and that values obtained with the direct determination technique and published prediction equations would differ. Thus, the objective of the present study was to determine the NE content of CM derived from Brassica napus yellow and Brassica juncea yellow using the direct determination technique (i.e. indirect calorimetry) in growing pigs, and to compare the NE values obtained with the direct determination technique and published prediction equations.
MATERIALS AND METHODS The experimental protocol used in the present study was reviewed and approved by the Animal Care Committee of the University of Manitoba. Animals were cared for according to the guidelines of the Canadian Council on Animal Care (CCAC 2009).
Animals A total of 18 growing barrows (Large White × Landrace × Duroc) with an average initial body weight (BW) of 14.9 ± 0.11 kg (mean ± SEM) were acquired from the Gleanlea Swine Research Unit, University of Manitoba. Pigs were individually housed for 15 days in adjustable metabolism crates (1.80 × 0.6 m) with smooth transparent plastic sides and plastic-covered expanded metal sheet flooring in a temperature-controlled room (22 ± 2°C).
Diets The basal diet was formulated based on corn-soybean meal to meet NRC (1998) requirements for growing pigs. Two test diets were formulated with 700 g/kg of the basal diet and 300 g/kg of CM (i.e. Brassica napus yellow or Brassica juncea yellow; Table 1).
Experimental design and procedures This study was conducted in two consecutive periods (10 pigs for period 1 and 8 pigs for period 2) using the same facility and similar experimental conditions and procedures because only two indirect calorimetry chambers were available at the time for the current study. Pigs were assigned to one of three experimental diets in a completely randomized design to give six replicates per diet. In each period, pigs were housed in metabolism crates and fed experimental diets for 15 days, © 2014 Japanese Society of Animal Science
Table 1 Composition and analysis of experimental diet (as-fed basis)
Item
Diets
Ingredients, g/kg Corn Soybean meal Brassica napus yellow Brassica juncea yellow Vegetable oil Salt Monocalcium phosphate Limestone Vitamin/mineral premix† Lysine-HCl DL-methionine L-threonine Calculated composition Crude protein, g/kg DE, MJ/kg ME, MJ/kg NE, MJ/kg SID lysine, g/MJ DE NDF, g/kg Crude fat, g/kg Analysed composition Crude protein, g/kg Dry matter, g/kg GE, MJ/kg Ash, g/kg NDF, g/kg ADF, g/kg Ether extract, g/kg Starch, g/kg
Basal
BNY
BJY
679.1 272.5
475.2 190.8 300.0
475.2 190.8
12.5 5.0 9.6 10.5 10.0 0.60 0.10 0.10
8.8 3.5 6.7 7.4 7.0 0.40 0.10 0.10
300.0 8.8 3.5 6.7 7.4 7.0 0.40 0.10 0.10
177 14.5 13.8 10.0 0.60 101 31
230 13.8 13.2 9.7 0.74 147 27
242 14.3 13.6 9.9 0.73 119 29
174 883 16.3 51 118 18 39 386
231 890 16.6 54 199 23 34 280
248 894 16.9 56 168 28 32 290
†Provided the following nutrients (per kg of air-dry diet): vitamins: A, 2000 IU, D3 200 IU, E, 40 mg, K, 2 mg, B1, 1.5 mg, B2, 7 mg, B6, 2.5 mg, B12, 25 μg, calcium pantothenate, 14 mg, folic acid, 1 mg, niacin, 21 mg, biotin, 70 μg. Minerals: Cu, 10 mg (as copper sulphate), iodine, 0.4 mg (as potassium iodine), iron, 120 mg (as ferrous sulphate), Mn, 10 mg (as manganous oxide), Se, 0.3 mg (as sodium selenite), Zn, 110 mg (as zinc oxide). BNY, Brassica napus yellow; BJY, Brassica juncea yellow; ADF, acid detergent fibre; DE, digestible energy; GE, gross energy; ME, metabolisable energy; NDF, neutral detergent fibre; NE, net energy; SID, standard ileal digestible.
including 10 days for adaptation to feed and environmental conditions and 5 days for total collection of feces and urine to measure DE and ME contents. Pigs were fed their respective diets at 2.3 MJ of ME/kg BW0.6 per day based on BW on days 1, 5 and 10 which was close to ad libitum intake (after Noblet et al. 1994b). During the study, pigs were fed at 08.30 hours and trained to consume their daily feed allowance within 1 h. Pigs had unlimited access to water via a low-pressure nipple throughout the study. During the last 5 days of each feeding period, total fecal and urine collection for the estimation of DE and ME was performed as previously described (Woyengo et al. 2010). On day 16, two pigs were transferred to the indirect calorimetry chambers (1.22 × 0.61 × 0.91 m; Columbus Instruments, Columbus, OH, USA) for 36 h of heat production (HP) and fasting HP (FHP) measurement based on O2 consumption, CO2 production and urine output. The following sets of two pigs were Animal Science Journal (2014) 85, 751–756
NET ENERGY CONTENT OF CANOLA MEAL
transferred to the indirect calorimetry chambers every 2 days (i.e. days 18, 20, 22, 24 and 26). Pigs were brought into the indirect calorimetry chambers within 1 h after consumption of their daily ration and HP was measured continuously for 24 h followed by 12 h of FHP measurement. Pigs were allotted to one of two indirect calroimetry chambers in a completely randomized design to avoid possible confounding effects. Fresh water was available in the indirect calorimetry chambers at all times and urine voided during the 24 h and 12 h period were collected separately, weighed, sub-sampled and stored at −20°C until required for N analysis. Temperature within the indirect calorimetry chambers was maintained at 22 ± 1°C and personnel movement around the indirect calorimetry chambers was limited to avoid disturbing pigs during HP and FHP measurements.
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The NE contents of the basal and CM containing diets are calculated according to the equations established by Noblet et al. (1994b):
NE (MJ /kg DM) = 0.843 × DE (MJ /kg DM) − 463
(1)
NE (MJ/kg DM) = 0.703 × DE (MJ /kg DM) + 1.58 × ether extract (% DM) + 0.47 × starch (% DM) (2) − 0.97 × crude protein (% DM) − 0.98 × crude fiber (%DM) NE (MJ /kg DM) = 0.700 × DE (MJ /kg DM) + 1.61 × ether extract (% DM) + 0..48 × starch (% DM) − 0.91 × crude (3) protein (% DM) − 0.87 × acid deterrgent fiber (% DM) The NE of test CM was calculated as follows:
Sample preparation and chemical analyses Dry matter (DM) was determined according to AOAC (1990; method 925.09) and gross energy (GE) content was measured using an adiabatic bomb calorimeter (model 6300; Parr Instrument, Moline, IL, USA), which had been calibrated using benzoic acid as a standard. N content was determined using the combustion method (990.03; AOAC 1990) using the LECO (model CNS-2000; LECO Corp., St. Joseph, MI, USA) N analyzer. Acid detergent fiber (ADF) and neutral detergent fiber (NDF) contents were determined according to the method of Goering and Van Soest (1970) and ash content was determined according to AOAC (1990; method 942.05). Ether extract (EE) was determined according to AOAC (1990; method 920.39). Starch content was measured according to the AOAC (1990; method 996.11) using an assay kit (Megazyme Total Starch assay kit; Megazyme International Ltd, Wicklow, Ireland). Non-starch polysaccharides were analyzed using gas-liquid chromatography (component neutral sugars) using an SP-2340 column and a Varian CP3380 gas chromatograph (Varian Inc., Palo Alto, CA, USA) and by colorimetry (uronic acids) using a Biochrom Ultrospec 50 (Biochrom Ltd, Cambridge, England) and the procedure described by Englyst and Cummings (1988) with some modifications (Slominski & Campbell 1990). The urine samples collected from the metabolism crates were analyzed for DM and GE. For the DM of urine, 1 mL of each sample was mixed with 0.5 g of cellulose and the weight of the resulting mixture recorded. The urine-cellulose mixtures together with samples of pure cellulose were dried in an oven at 50°C for 24 h. The GE was then determined on the dried urine-cellulose mixtures as described above and samples of pure cellulose, and the contents of the same in urine were calculated by the difference method (Fleischer et al. 1981).
Calculations Heat production, FHP, retained energy (RE) and NE were calculated using the following equations:
HP (MJ/kg DM) = [3.87 × O2(L) + 1.20 × CO2(L) − 1.43 × urinary N (g)]/dry matter intake (DMI)
(Brouwer 1965) RE (MJ /day) = [ME (MJ/day) − HP (MJ /day)]/DMI NE (MJ /kg DM) = (RE MJ /day + FHP MJ /day)/DMI Animal Science Journal (2014) 85, 751–756
NETest canola meal(MJ/kg DM) = Basal diet NE − Basal diet NE − Test diet NE/0.3
Statistical methods Treatment effects were evaluated univariately in a normal mixed-liner model using the GLM procedures of SPSS (version 18.0, SPSS Inc., Chicago, IL, USA). The pig was considered as the experimental unit for all measurements and the model included dietary treatments. The experimental period was used as a random factor. Statistical significance was accepted at P < 0.05.
RESULTS All animals adapted well to their respective diets and environmental conditions, remained healthy and readily consumed their daily feed allowance during the experimental period. Nitrogen balance and HP and FHP values obtained with the direct determination technique are presented in Table 2. Nitrogen retention for pigs fed Brassica napus yellow- and Brassica juncea yellow-containing diets were 19.4 and 22.0 g/day, respectively. Corresponding values for HP were 7.3 and 7.8 MJ/kg, respectively. The energy balance of pigs and energy values of experimental diets and CM determined with the direct determination technique and prediction equations are presented in Table 3. Using the direct determination technique, the DE, ME and NE contents in Brassica napus yellow and Brassica juncea yellow were 13.6, 12.6 and 8.8 and 14.2, 13.5 and 9.8 MJ/kg DM, respectively. Respective values obtained with published prediction equations were 13.8, 12.8 and 8.1 and 15.0, 13.9 and 9.4 MJ/kg DM for Brassica napus yellow and Brassica juncea yellow, respectively. The RE and FHP of Brassica napus yellow- and Brassica juncea yellow-containing diets were 5.5 versus 5.7 MJ/kg and 4.3 versus 4.5 MJ/kg, respectively. The NE contents of diets containing Brassica napus yellow and Brassica juncea yellow were 9.8 and 10.2 MJ/kg DM and the NE contents of Brassica napus yellow and © 2014 Japanese Society of Animal Science
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Table 2 Nitrogen balance in pigs fed diets containing canola meals determined with direct determination technique
Item
Nitrogen balance Nitrogen ingested, g/day Nitrogen output in faces, g/day Nitrogen digested, % Nitrogen retained, g/day Urine nitrogen excretion, g/day
Diets Basal
Brassica napus yellow
Brassica juncea yellow
18.3 4.3 76.6 14.0 4.8
27.0 7.6 71.8 19.4 4.3
29.1 7.2 75.3 22.0 4.7
SEM
P-value
0.33 0.19 0.70 0.27 0.42
0.001 0.001 0.036 0.001 0.878
Table 3 Energy balance of pigs and energy values of experimental diets and canola meals determined with direct determination technique using the indirect calorimeter chamber and prediction equations using chemical characteristics
Item
Direct determination technique DE of diets, MJ/kg DM DE of canola meal, MJ/kg DM ME of diets, MJ/kg DM ME of canola meal, MJ/kg DM Heat production, MJ/kg Fasting heat production, MJ/kg Retained energy, MJ/kg NE of diets, MJ/kg DM NE of canola meal, MJ/kg DM Prediction equations provisions DE of diets, MJ/kg DM† DE of canola meal, MJ/kg DM† ME of diets, MJ/kg DM2 ME of canola meal, MJ/kg DM‡ NE of diets, MJ/kg DM§ NE of canola meal, MJ/kg DM§
Diets
SEM
P-value
Basal
Brassica napus yellow
Brassica juncea yellow
15.5 – 15.1 – 8.4 4.5 6.1 10.6 –
14.8 13.6 14.3 12.6 7.3 4.3 5.5 9.8 8.8
14.9 14.2 14.2 13.5 7.8 4.5 5.7 10.2 9.8
0.10 0.35 0.10 0.31 0.18 0.90 0.29 0.25 0.58
0.021 0.458 0.004 0.213 0.091 0.826 0.645 0.484 0.466
15.0 – 14.6 – 11.0 –
14.7 13.8 14.2 12.8 10.3 8.1
15.1 15.0 14.5 13.9 10.4 9.4
0.79 0.89 0.04 0.06 0.62 0.23
0.980 0.546 0.029 0.001 0.007 0.052
†Mean of prediction equations by Noblet and Perez (1993), where DE = 949 + (0.789 × GE) – (43 × % Ash) – (41 × % NDF) and DE = 4151 – (122 × % Ash) + (23 × % CP) + (38 × % EE) – (64 × % CF). ‡Mean of prediction equations by May and Bell (1971) and Noblet et al. (1989) and Noblet and Perez (1993), where ME = DE × [1.012 – (0.0019 × % CP)], ME = DE × [0.998 – (0.002 × % CP)] and ME = DE × [1.003 – (0.0021 × % CP)], respectively. §Mean of prediction equations by Noblet et al. (1994a), where NE = 0.843 × DE – 463, NE = 0.703 × DE + 1.58 × (%) EE + 0.47 × (%) starch – 0.97 × (%) CP – 0.98 × (%) CF and NE = 0.700 × DE + 1.61 × (%) EE + 0.48 × (%) starch – 0.91 × (%) CP − 0.87 × (%) ADF. BW, body weight; DE, digestible energy; ME, metabolisable energy; SEM, pooled standard error of mean; GE, gross energy; NDF, neutral detergent fiber; CP, crude protein; EE, ether extract; CF, crude fiber; ADF, acid-detergent fiber.
Brassica juncea yellow were determined to be 8.8 and 9.8 MJ/kg DM, respectively, by the direct determination technique. However, the average NE contents of Brassica napus yellow and Brassica juncea yellow determined using published prediction equations were 8.1 and 9.4 MJ/kg DM, respectively.
DISCUSSION In the present study, we have carefully selected the three prediction equations based on the DE system because the DE values are extensively available and could be effortlessly found for most feed ingredients, particularly in North America, Asia and Australasia and these equations were highly correlated (r2 = 0.910.97). In this light, we have used the mean NE values obtained from these prediction equations to compare © 2014 Japanese Society of Animal Science
with values obtained from the direct determination technique. The calculated NE content of experimental diets used in the present study were closer to the values obtained using the direct determination technique than values obtained using prediction equations reported by Noblet et al. (1994b) (see Tables 1 and 3). Nevertheless, the calculated contents of DE and ME were higher compared to the values obtained from either of the two techniques used in the current study. This discrepancy could be due to the high protein and fiber contents of experimental diets which are known to overestimate energy values when expressed on a DE and ME system compared with energy values measured in feed containing high contents of starch and fat in pigs (Noblet et al. 1994a). The analyzed gross energy, ash, EE and NDF content of test ingredients (i.e. Brassica napus yellow and Brassica juncea yellow) Animal Science Journal (2014) 85, 751–756
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were similar to reported values (e.g. Slominski et al. 1999; Montoya & Leterme 2009), but the crude protein content of the Brassica napus yellow was lower compared with values (393 vs. 466 and 519 g/kg DM, respectively) reported in the above cited studies. We suspected that this response could be attributed to excessive toasting of Brassica napus yellow because it has been reported that increasing the duration of toasting had a negative impact on soluble protein content (Barac´ & Stanojevic´ 2005). Likewise, van Barneveld et al. (1994) reported that dietary inclusion of heat-treated ingredients had significant reductions in protein deposition, ileal-digestible lysine retention and growth performance in growing pigs. This could have contributed to the relatively lower NE content of Brassica napus yellow independent of the determination techniques (i.e. direct determination vs. prediction equation; 8.8 vs. 8.1 MJ/kg DM, respectively) compared with a value of 11.9 MJ/kg DM for the same canola meal type published elsewhere (Montoya & Leterme 2009). However, the NE contents of the Brassica juncea yellow determined in this study using both techniques (i.e. direct determination vs. prediction equation; 9.8 vs. 9.4 MJ/kg DM) were comparable with values of 9.4 MJ/kg DM reported by Montoya and Leterme (2009). In addition, the NE content of Brassica juncea yellow is calculated to be 9.6 MJ/kg DM when the same prediction equation was applied with the cited study (i.e. equation 3); however, it was not possible in the present study to reconcile the accuracy between published equations used as these were developed using complete diets with various inclusion levels of ingredients. In the present study, the NE content determined using the direct determination technique was approximately 5.9% higher than the value calculated using the prediction equations (Noblet et al. 1994b). It is known that energy utilization is greater in mature pigs (Noblet & Shi 1993; Le Goff & Noblet 2001), and abnormality is essential when applying this theory because pigs used in the present study were lighter (about 26 kg) than those used (about 35–45 kg) in the studies of Noblet et al. (1994a, b). However, feeding levels, digestive physiological responses and digestive utilization of diets in pigs at around 25 to 50 kg would not relatively differ (NRC 2012). Furthermore, it is likely that the NE values of diets and ingredients could differ due to genetics, methodologies used to measure RE, feeding strategies and environmental conditions in which pigs are kept (Boisen & Verstegen 1998). In addition, the discrepancy could be due to those prediction equations per se because these were derived using complete diets. The NE content in Brassica napus yellow was lower compared with Brassica juncea yellow regardless of the NE determination technique used. This could be attributed to the lower crude protein and starch content but Animal Science Journal (2014) 85, 751–756
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Table 4 Analysed chemical composition of ingredients (g/kg, as-fed)
Item
Brassica napus yellow
Brassica juncea yellow
Crude protein Dry matter GE, MJ/kg Ash NDF ADF Crude fat Ether extract Starch Non-phytate phosphorus Total NSP
355 903 17.8 73 253 64 82 21 4 4.0 217
394 901 17.8 70 159 58 69 23 22 3.9 250
ADF = acid detergent fibre; GE = gross energy; NDF = neutral detergent fibre; NSP = non-starch polysaccharides.
higher NDF content of Brassica napus yellow than in Brassica juncea yellow (Table 4). It is known that dietary fiber (e.g. non-starch polysaccharides and NDF) is less digestible (< 50%) than other nutrients (e.g. crude protein and starch: 80–100%; Noblet & van Milgen 2004). Likewise, nitrogen balance was significantly lower in Brassica napus yellow compared to pigs fed diet containing Brassica juncea yellow that also could have contributed to lower NE content in Brassica napus yellow compared with pigs fed diet containing Brassica juncea yellow (Table 2). Nevertheless, we have used different levels of vegetable oil content in the experimental diets because only basal diet included vegetable oil. Nonetheless, Kil et al. (2011) demonstrated that the NE content of a corn-soybean meal containing 5% of soybean oil and the counterpart containing 10% of soybean oil were not affected by the content of dietary lipids (2206 vs. 2318 kcal/kg, respectively).
Conclusions The NE contents of Brassica napus yellow and Brassica juncea yellow were 8.8 versus 9.8 and 8.1 versus 9.4 MJ/kg between the direct determination technique using the indirect calorimetry chamber and prediction equations using chemical characteristics, respectively. This study showed that there is discrepancy of approximately 5.9% in the NE content between the NE determination techniques, suggesting that great variation in the NE content of diets and ingredients could be ascribed to inherent factors (i.e. BW, gender and feeding level) in relation to using different methodologies. Furthermore, research is warranted to examine the effect of the NE evaluated between the direct determination technique and prediction equations on growth performance in various BW ranges of pigs. © 2014 Japanese Society of Animal Science
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ACKNOWLEDGMENTS The authors thank Mr. Robert Stuski, Mr. Darwin Ramos, Mr. Atanas Karamanov and Ms. Lisa Rigaux for technical assistance. This research was supported by Canola Council of Canada through the Canola Science Cluster.
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