Microsoft Word - 13-Agra_33105 113 Original Article Biosci. J., Uberlândia, v. 33, n. 1, p. 113-120, Jan./Feb. 2017 THE CRAMBE (Crambe abyssinica Hochst) BYPRODUCTS, CAN BE USED AS A SOURCE OF NON-DEGRADABLE PROTEIN IN THE RUMEN? COPRODUTOS DE CRAMBE (Crambe abyssinica Hochst) PODEM SER UTILIZADOS COMO FONTE DE PROTEINA NÃO DEGRÁDAVEL NO RÚMEN? Rafael Henrique de Tonissi e Buschinelli de GOES1; Rosiélen Augusto PATUSSI2; Jefferson Rodrigues GANDRA1; Antonio Ferriani BRANCO3; Thiago José de Lira CARDOSO2; Marcus Vinícius Moraes de OLIVEIRA4; Raquel Tenório de OLIVEIRA5; Charles Jhonnatan dos Santos SOUZA5 1. Professor, Doutor, Faculdade de Ciências Agrárias. Universidade Federal da Grande Dourados - UFGD, Dourados, MS, Brasil. rafaelgoes@ufgd.edu.br; 2. Mestre em Zootecnia, Programa de Pós-Graduação em Zootecnia, UFGD, Dourados, MS. Brasil; 3. Professor, Doutor, Departamento de Zootecnia – DZO, Universidade Estadual de Maringá, Maringá, PR, Brasil; 4. Professor, Doutor, Universidade Estadual de Mato Grosso do Sul – UEMS, Aquidauana, MS, Brasil; 5. Estudante de Graduação em Zootecnia, UFGD, Dourados, MS, Brasil. ABSTRACT: To evaluated the chemical composition and ruminal degradability of crambe byproducts (meal and crushed) and proteic supplements formulated with crushed crambe (0; 2.5; 5; 10 and 15%); five crossbred steers with average weight of 485±14 kg, were used. All the animal were kept in individual paddocks of 0.25 ha on Urochloa brizantha (syn. Brachiaria brizantha). It was observed a greater soluble fraction, higher effective degradability at 5%/h and higher degradation rate “c” and, consequent, lower indigestible fraction for crambe crushed ground in the sieve of 3 mm. The effective degradability at 5%/h was lower for the crambe crushed (55.42) in relation to the meal (48.80). The diet with 5% of inclusion of crambe showed higher effective degradability for dry matter (54.86%) and lower fraction “I” (30.64%) associated with higher fractions “c” and “b”. Crambe ground in sieves of 1 and 3 mm mesh presented the highest degradability. Crushed crambe showed higher ruminal degradation than crambe meal; the crambe byproducts possibility can be use as a source of non-degradable protein in the rumen. KEYWORDS: Crambe meal. Crushed crambe. In situ degradability. INTRODUCTION Oilseeds are used in ruminant diets due to their high concentrations of lipids, and composition of fatty acids, rich in unsaturated fatty acids (ω-3 and ω-6), and because they present a slow release of oil, due to chewing, which results in small fractions coming into the rumen (CIEŚLAK et al., 2013). Crambe (Crambe abyssinica Hochst) has been widely used for the extraction of oil for biodiesel production. Vegetable oils used for biodiesel production can be extracted from oilseeds through two different processes: with the use of solvents or by pressing. The waste from the use of solvents for oil extraction is meal and from pressing, crushed grain. The seed has between 46 and 58% CP with 44% EE (Souza et al. 2009; Goes et al., 2010). Crambe meal contains about 47.4 µ mol/g of glucosinolates [(S)-2-hydroxy-3-butenyl glucosinolate]; and it´s toxic (11-24,3 mol/g) to many organisms and impairs the activity of rumen flora in cattle after six days of ingestion (TRIPATHI; MISHRA, 2007). Considering the high nutritional potential of this oilseed, and if crambe products can be used in ruminant nutrition, the present study evaluated by the in situ technic the kinetic patterns of ruminal degradation of dry matter and crude protein of crambe byproducts, and, degradability of supplements with inclusion of crushed crambe replacing soybean meal. MATERIAL AND METHODS The experiments were conducted at the Sector of Ruminant Nutrition and Animal Nutrition Laboratory of the Federal University of Grande Dourados. In both experiments we used, crossbred steers with average weight of 485±14 kg, cannulated in the rumen were kept in individual paddocks (0.25 ha) of Urochloa brizantha (syn. Brachiaria brizantha) cv. Marandu (7047.85 kg of DM/ha); receiving daily, in the morning (8:00h), 8 g/kg body weight of supplement containing crushed crambe (Table 1). Experiment 1 Crambe byproducts (crushed and meal) were evaluated in two trials; on the first trial we evaluated the ruminal degradability of crambe Received: 04/02/16 Accepted: 05/12/16 114 The crambe (Crambe abyssinica Hochst) byproducts… GOES, R. H. T. B. et al. Biosci. J., Uberlândia, v. 33, n. 1, p. 113-120, Jan./Feb. 2017 crushed at different particle size (1, 3 and 5 mm) and the second trial were evaluated the ruminal degradability of crambe meal and crambe crushed grounded at 3mm. The chemical composition of crambe byproducts presents in Table 2. In both trials the crushed crambe was obtained by cold pressing. Table 1. Percentual to the experimental supplements. Ingredients (g/kg) C0 C2.5 C5 C10 C15 Crushed crambe - 25 50 100 150 Soybean meal 150 125 100 50 - Rice meal 400 400 400 400 400 Corn meal 376 375 373 369 366 Urea 3.5 5.2 7.0 10.5 14 Salt 10 10 10 10 10 Limestone 25 25 25 25 25 Sulfur 10 10 10 10 10 Dicalcium phosphate 15 15 15 15 15 Premix2 10 10 10 10 10 1C00=supplement without crambe crushed; C2.5= supplement with 2.5% crambe crushed; C5= supplement with 5% crambe crushed; C10= supplement with 10% crambe crushed; C15= supplement with 15% crambe crushed.2Calcium: 120.00 g. phosphorus: 88.00 g. iodine: 75.00 mg. manganese: 1300.00 mg. sodium: 126.00 g. selenium: 15.00 mg. sulfur: 12.00 mg. zinc: 3630.00 mg. cobalt: 55.50 mg. cooper: 1530.00 mg e iron: 1800.00 mg. Table 2. Chemical composition (g/kg) and in vitro dry matter digestibility (IVDMD) (%) of crambe byproducts. Feedstuffs DM OM CP EE NDF ADF LIG IVDMD Crushed Crambe 943.0 952.2 261.9 182.7 302.3 194.4 84.0 62,04 Crambe Meal 899.0 931.9 350.0 41.0 350.0 242.0 113.0 58,61 DM = Dry matter, OM = organic matter, CP = crude protein, EE=ether extract, NDF = neutral detergent fiber, ADF = acid detergent fiber, MM = mineral matter, LIG = lignin. The feedstuffs were analyzed to dry matter (DM – method 930.15); ash (Ash - method 942.05) and the organic matter (OM = 100 - ash); crude protein (CP – method 976.05, N X 6.25) and ether extract (EE – method 920.39), following the methodologies of AOAC (2006). ADF contents were obtained following method described by Van Soest and Robertson (1985). Lignin content was obtained by oxidation with potassium permanganate (VAN SOEST; WINE, 1968). For analysis of NDF, samples were treated with heat stable alpha amylase without sodium sulfite and corrected for ash residue (MERTENS, 2002). The feedstuffs were dried at 65ºC for 24 hours, removed, and weighed. After weighing, the food was packed in TNT bags (TNT -100 g/m2) in size 5.0 x 5.0 cm, respecting the relationship 20 mg / cm² (CASALI et al. 2009). The samples were prepared and incubated according to Nocek (1988) and Huntington & Givens (1995). TNT Bags were introduced directly into the rumen, in decreasing order of 48, 36, 24, 12, 8, 4, 2 and 0 hours, in triplicates per animal/incubation time, according to NRC (2001); and . removed all at once and rinsed in tap water, until clean. The remaining residues from the incubation were oven dried at 65ºC for 48h and stored for later analysis to determine the variables studied. The disappearance of dry matter and crude protein (N x 6.25) were based on the weight difference between the incubated material and the residues after incubation. To estimate the kinetic parameters of DM and CP, we used the first-order asymptotic model described of Ørskov e McDonald (1979): PD = a+b(1-e-ct). Where: PD=potential rumen degradability; a=soluble fraction; b=potentially degradable fraction of the insoluble fraction that would be degraded at a rate c; c=degradation rate of the fraction “b”; t=incubation time in hours. The fraction considered as undegradable is calculated according to Ørskov & McDonald (1979): I = 100-(a+b). And the effective degradability (ED) is calculated with the following equation: ED = a + [(b*c)/(c+K)]; where K=passage rate of solids from the rumen, herein defined as 2, 5 and 8.0% per hour (h), which can be attributed to the low, medium and high dietary intake. After fitting the data to the model and using the disappearance value obtained at the time zero (a’), we estimated the colonization time (CT) as proposed by Patiño et al. (2001), where the 115 The crambe (Crambe abyssinica Hochst) byproducts… GOES, R. H. T. B. et al. Biosci. J., Uberlândia, v. 33, n. 1, p. 113-120, Jan./Feb. 2017 parameters a, b and c were estimated by the Gauss Newton algorithm: TC = [-ln(a’-a-b)/c]. Experiment 2 The inclusion levels of crambe crushed in the concentrates was 0; 2.5; 5.0; 10 and 15% (Table 1). Three samples were taken of each concentrate, which were placed in plastic bags, identified and stored at -20°C. For in situ degradability, the protein supplements were ground in 3 mm sieve, and then dried at 65ºC for 24 hours and weighed. Bags were introduced directly into the rumen, in descending order of 48, 36, 24, 12, 8, 4, 2 and 0 hours, in triplicate per animal/incubation time, according to NRC (2001). Each supplement was incubated in an animal receiving the same treatment. The rest of the methodologies used in the in situ degradability trial of protein supplements were similar to those in the experiment 1. Statistical analysis The animals were distributed in a randomized latin square design (5x5); and the degradation curves of dry matter and crude protein of feedstuffs evaluated, for each animal used, were subjected to fit by the respective models using the PROC NLIN of SAS 9.2. The in vitro dry matter digestibility was analyzed by polynomial regression PROG REG of SAS 9.2, with significance level of 5%. RESULTS Experiment 1 - trial 1 The kinetic parameters of in situ degradation for different particle sizes of crushed crambe were similar, with intermediate degradability for DM and CP (Table 3). Table 3. Kinetic parameters of in situ degradation of crushed crambe in different particle sizes Parameters* Effective degradability(%.h-1) Dry matter a (%) b (%) c(%/h) I (%) 2 5 8 r2 CT (min) 1 mm 27.86 22.24 0.08 49.90 45.31 41.16 38.65 88.75 342 3 mm 28.16 27.01 0.17 44.82 47.83 43.21 40.80 97.04 346 5 mm 26.20 23.37 0.07 50.42 44.11 39.46 36.73 91.49 353 Crude protein 1 mm 25.64 36.89 0.25 37.47 59.71 56.24 53.43 90.90 301 3 mm 31.55 28.56 0.31 39.89 58.36 56.11 54.21 88.28 271 5 mm 34.33 28.67 0.19 37.00 59.15 55.42 52.88 74.27 314 *a=soluble fraction; b= potentially degradable fraction; c= degradation rate of fraction b. I=indigestible fraction. CT = Time of colonization (minutes) It was observed a greater soluble fraction, higher effective degradability at 5%.h-1 and degradation rate “c”, consequent, lower indigestible fraction for feedstuffs grounded of 3 mm (28.16%, 43.21% and 44.82%, respectively). The crushed crambe grounded of 3 mm showed the highest degradation rates, and greater potentially degradable fraction of DM. For CP, the ground in the 5 mm demonstrated the highest soluble fraction, which did not result in higher effective degradability. The soluble fraction observed for the ground of 5 mm was 34.33%, but this value not correspond a greater effective degradability at 5%.h-1 (55.42%), lower than that found for the sieves of 1 and 3 mm. Experiment 1 - trial 2 The kinetic parameters of in situ degradability, there was a higher percentage for the fraction “a” and fraction “I” for the DM of the crushed crambe (Table 4). In turn, the meal showed higher percentage of the fractions “b” and “c” (%.h- 1). Also, we observed a low degradability at 5%.h-1 for both byproducts. The crushed presents a higher value for fraction “a” and “c”, by CP, and the meal have a higher percentage of the fractions “b”. The effective degradability at 5%.h-1 was lower for the crushed crambe (55.42%) in relation to the meal (48.80%). Experiment 2 The crushes crambe reduces the NDF of supplements, but increases the EE (Table 5). The diet with 5% of inclusion showed higher effective degradability for dry matter (54.86%) and lower fraction “I” (30.64%) associated with higher fractions “c” and “b”. The lower soluble fraction “a” and lower effective degradability at 5%.h-1, were verified in the 10% of 116 The crambe (Crambe abyssinica Hochst) byproducts… GOES, R. H. T. B. et al. Biosci. J., Uberlândia, v. 33, n. 1, p. 113-120, Jan./Feb. 2017 inclusion, with 6.68 and 49.08% respectively for DM (Table 6). Table 4. Kinetic parameters of in situ degradation crambe byproducts. Parameters* Effective degrabability (%.h-1) Dry matter a (%) b (%) c(%/h) I (%) 2 5 8 r2 CT (min) Crushed 26.20 23.17 0.07 50.63 43.96 39.35 36.64 91.49 353 Meal 18.94 31.39 0.11 49.67 29.17 38.26 34.95 99.14 352 Crude protein Crushed 34.33 28.67 0.19 37.00 59.15 55.42 52.88 74.27 316 Meal 20.68 43.24 0.10 36.08 37.44 48.80 44.05 94.44 364 *a=soluble fraction; b= potentially degradable fraction; c= degradation rate of fraction b. I=indigestible fraction. CT= colonization time in minutes Table 5. Chemical composition of supplements contents crushed crambe Nutrients1 Levels of crushed crambe (%) Average MSE2 P