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Growth and carcass characteristics of Santa Inês lambs fed diet supplemented with physic nut meal free of phorbol ester
Small Ruminant Research 114 (2013) 20–25
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Growth and carcass characteristics of Santa Inês lambs fed diet supplemented with physic nut meal free of phorbol ester P.B. Oliveira a , P.M.T. Lima a,∗ , A. Campeche b , S. Mendonc¸a c , B.G. Laviola c , C. McManus a , H. Louvandini b a b c
Universidade de Brasília, Campus Universitário Darcy Ribeiro, 70910900 Brasília, DF, Brazil Centro de Energia Nuclear na Agricultura, Universidade de São Paulo, Av. Centenário, no 303, 13416000 Piracicaba, SP, Brazil Embrapa Agroenergia, Parque Estac¸ão Biológica, 70770901 Brasília, DF, Brazil
a r t i c l e
i n f o
Article history: Received 11 June 2012 Received in revised form 13 May 2013 Accepted 14 May 2013 Available online 10 June 2013 Keywords: Bio-fuel By-product Carcass Jatropha curcas
a b s t r a c t The aim of this work was to evaluate the growth performance, carcass traits, hematological and biochemical variables of Santa Inês lambs fed diet supplemented with physic nut (Jatropha curcas) meal (PNM). Twenty-four intact male lambs [age: 120 ± 0.98 days; body weight (BW): 19.7 ± 2.4 kg] were assigned randomly to four concentrate mixtures. These mixtures were supplemented with 0 (CON), 100 (PNM100), 200 (PNM200), or 300 (PNM300) g/kg dry matter (DM) of PNM. The concentrate was offered at 1.2% BW, whereas lambs were fed Tifton (Cyndon dactylon CV. Tifton-85) hay ad libitum. Animals were weighed and blood collected every 15 days. At the end of the experiment, lambs were slaughtered and their carcasses were evaluated. Biochemical and hematological variables were not different among diets (P > 0.05). Similarly, dry matter intake was not different among diets (P > 0.05) and no acceptability problems were observed for the PNM. Average daily gain and carcass characteristics also did not differ (P > 0.05) between the treatments. No clinical symptoms of intoxication by physic nut were detected during the whole experimental period. Therefore, it may be inferred that the non-toxic PNM was shown to be a promising protein source for sheep nutrition and can be used in concentrations of up to 300 g/kg DM in concentrate mixture. © 2013 Elsevier B.V. All rights reserved.
1. Introduction Agro-industry by-products have been used mainly to reduce production costs, and to offset increasing environmental concerns about less expensive waste management programs (Grasser et al., 1995). Studies have demonstrated that residues of the bio-fuel industry may have satisfactory levels of energy and protein, supplying requirements and
∗ Corresponding author at: Centro de Energia Nuclear na Agricultura, ´ no 303, 13416000, Piracicaba, Universidade de S˜ao Paulo, Av. Centenario, SP, Brazil. Tel.: +55 061 82784646; fax: +55 019 34294728. E-mail addresses: [email protected], [email protected] (P.M.T. Lima). 0921-4488/$ – see front matter © 2013 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.smallrumres.2013.05.007
substituting commonly used ingredients, such as soybean meal and corn grains. Among these residues, physic nut meal (PNM) is not widely used, as it is regarded as toxic for animals (Abdalla et al., 2008). It was previously believed that the toxicity of the physic nut is due to the presence of curcin, which has a toxic activity (Ribosome-Inactivating Protein – RIP) similar to the castor bean (Ricinus communis) ricin (Felix et al., 2008; Mendonca and Laviola, 2009). However, recent research has shown that the toxicity of physic nut seeds and oil is mainly due to the presence of phorbol ester, and not to curcin (Abdalla et al., 2008). One of the great advantages of physic nut is its production cycle, which may last for 40 years, maintaining an average productivity of two tons per hectare (Mendonca
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and Laviola, 2009). The physic nut seed has approximately 210 g/kg crude protein (CP) and 400 g/kg ether extract (EE), while PNM has 286 g/kg CP and 142 g/kg EE on dry matter basis (Souza et al., 2009). Presently, there are more than 30 thousand hectares of physic nut in Brazil, distributed over the Southeast, Center-West and Northeast regions of the country. Considering only the plants that have reached the mature stage, there is a potential for producing more than 90 thousand tons of seeds per year, which would correspond to 58.5 thousand tons of PNM per year (Mendonca and Laviola, 2009). With new detoxification techniques and varieties of physic nut with zero concentration of phorbol ester (nontoxic), new uses for these residues arise. This protein rich feed (detoxified or non-toxic), which has phorbol ester as its limiting factor, is still not well studied for use as a feedstuff in animal nutrition. Generally, pressedcakes or meals generated by the oil extraction process do not pass through any kind of value-adding procedure, and their economic and nutritional potential, except for some materials from soybean or cottonseed processing, are unknown (Abdalla et al., 2008; Mendonca and Laviola, 2009). The objective of this study was to evaluate growth performance, carcass traits, as well as hematological and serum biochemical variables of Santa Inês lambs fed different levels of non-toxic physic nut meal in the diet. 2. Materials and methods 2.1. Study area The experiment was carried out at the Sheep Management Center, in the Água Limpa Experimental Station of the University of Brasília. The animals were allocated in individual pens of 1.8 m2 each, disposed in a 240 m2 shed.
2.2. Animals and diets Twenty-four male Santa Inês lambs [age: 120 ± 0.98 days; body weight (BW): 19.7 ± 2.4 kg] were divided into four diets (6 lambs/diet). Diets were: (1) control (270 g/kg soybean meal and 730 g/kg ground corn; CON); (2) 220 g/kg soybean meal, 680 g/kg ground corn and 100 g/kg PNM; PNM100; (3) 180 g/kg soybean meal, 620 g/kg ground corn and 200 g/kg PNM; PNM200; and (4) 140 g/kg soybean meal, 560 g/kg ground corn, and 300 g/kg PNM; PNM300. The experimental period lasted for 70 days. The physic nut meal (PNM) that was used in the current study was considered not toxic because phorbol ester was not detected when analyzed according to procedure described by Makkar and Becker (1999). The PNM (Germplasm Bank of Embrapa Agroenergy – CNPAE-169 and CNPAE-170) was purchased from a rural property in Mato Grosso do Sul State and was provided for the experiment by Embrapa Agroenergy after oil extraction by mechanical press. Each lamb was allocated to an individual pen (1.60 m × 1.15 m). The diets were balanced to offer 180 g/kg of CP and 800 g/kg of total digestive nutrients. The PNM was included in each diet as a substitute of total protein. Lambs had ad libitum access to water and Tifton (Cyndon dactylon CV. Tifton-85) hay. The chemical composition of the hay was 872.0, 68.8, 759.0, 410.0, 7.9, 51.0, and 534 g/kg DM, CP, neutral detergent fiber (NDF), acid detergent fiber (ADF), ether extract (EE), ash and total digestible nutrients (TDN), respectively. The concentrate was offered at 1.2% BW. The forage and concentrate mixture were offered separately, so that the PNM intake could be precisely calculated. The chemical analyses of the Tifton hay and concentrates were performed according to AOAC (1995) and Mertens (2002).
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Table 1 Chemical composition of concentrates used in the diets. Nutrient (g/kg)
Dry matter Crude protein Neutral detergent fiber Acid detergent fiber Ether extract Ash Total digestible nutrientsb
Dietsa CON
PNM100
PNM200
PNM300
879.0 201.0 108.0 57.0 35.1 19.9 810.0
883.0 200.8 140.0 70.2 38.9 18.7 808.0
887.0 200.5 171.4 88.4 42.1 17.9 807.0
890.0 200.3 200.1 106.0 45.5 17.2 807.0
a Control (CON) = 0 g/kg physic nut meal (PNM), 100 g/kg PNM (PNM100), 200 g/kg PNM (PNM200), and 300 g/kg PNM (PNM300). b Calculated according to Harlan et al. (1991).
To calculate dry matter intake (DMI), the leftovers of forage were collected and weighed three times a week. There were no concentrate leftovers. Lambs were weighed fortnightly, before they received the concentrate in the morning, throughout experimental period. After weighing, blood was collected via jugular venipuncture using vacuum tubes with and without anticoagulant (EDTA). The hemogram analyses were performed using an ABX – model 22P ABX micro 60® automatic counter. Biochemical tests (aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), creatinine and albumin) were performed using blood serum and analyses were carried out using commercial biochemical tests kits Labtest® and spectrophotometer Bioplus, model Bio-2000® . On the last day of the experimental period, the animals were slaughtered so that the carcass characteristics could be evaluated, according to procedures described by Osório et al. (1998). The slaughtering took place after a 16 h fast, following the current regulations in Brazil, Regulation of Industrial and Sanitary Inspection for Products of Animal Origin – RIISPOA (1952). Immediately before the slaughtering, animals were weighed to obtain fasting body weight (FBW). The animals were stunned by electrocution. After bleeding, skinning and evisceration, the viscera were weighed in two groups: Group A (rumen, reticulum, omasum, abomasum and intestines) and Group B (tongue, esophagus, trachea, lungs, heart, liver and kidneys). During slaughter, scrotal circumference was also measured. The whole animal carcass was used to determine hot carcass weight (HCW), and hot carcass yield (HCY), using the equation: HCY = HCW/FBW × 100. Carcasses were cooled in cold chamber, at temperatures below 4 ◦ C + 2 ◦ C for 24 h and weighed again to obtain cold carcass weight (CCW). Cold carcass yield (CCY) was calculated: CCY = CCW/FBW × 100 and the cooling loss index, CLI = (HCW − CCW)/HCW × 100. A longitudinal cut was made along the vertebral column, splitting the carcass in two parts, which were then weighed. The carcass was cut into the following parts: neck, shoulder, loin, ribs, flank steak, and leg which were weighed individually. 2.3. Statistical analysis The experiment was conducted in a completely randomized design. The data analysis was carried out by the software SAS v.9.2® (Cary, North Carolina), using the MIXED procedure for measures repeated over time. For the variables measured only once, the GLM procedure was employed using ANOVA analysis. The REG procedure was used to study the effects of increasing PMN in the diet over all the variables evaluated. For all the analysis, the significance level adopted was 5% (P < 0.05).
3. Results The chemical composition of concentrates used in the diet is presented in Table 1. Growth performance of Santa Inês lambs fed diets with or without physic nut meal is shown in Table 2. No significant differences (P > 0.05) were observed in DMI, average daily gain (ADG), and feed conversion ratio between diets.
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Table 2 Growth performance of Santa Inês lambs fed diets containing physic nut meal. Dietsa
Variables
Duration (days) Initial BWc (kg) Final BW (kg) ADG (g/animal/day) DMI (g/animal/day) FCR (g gain/kg feed) a b c d e
CON
PNM100
PNM200
PNM300
SDb
70 19.6d 29.4 139.3 1053.8 132.2
70 19.5 28.2 123.8 1002.1 126.4
70 19.8 28.6 126.2 1013.8 124.5
70 19.8 28.5 123.8 1010.7 122.5
– 2.4 2.5 22.2 214.2 21.17
Regression Linear
Quadratic
– NSe NS NS NS NS
– NS NS NS NS NS
Control (CON) = 0 g/kg physic nut meal (PNM), 100 g/kg PNM (PNM100), 200 g/kg PNM (PNM200), or 300 g/kg PNM (PNM300). SD, standard deviation. BW, body weight; ADG, average daily gain; DMI, dry matter intake; FCR, feed conversion ratio. The means presented here were not significantly different (P > 0.05) among diets. NS, not significant (P > 0.05).
Table 3 Carcass traits of Santa Inês lambs fed diets with or without physic nut meal. Dietsa CON
PNM100
PNM200
PNM300
SDb
Regression Linear
c
FBW (kg) HCW (kg) CCW (kg) HCY (%) CCY (%) CLI (%) a b c d e
d
26.7 10.6 10.2 39.9 38.3 3.7
26.6 11.0 10.6 41.7 40.3 3.2
27.3 11.2 10.8 41.0 39.6 3.0
25.2 11.5 11.1 45.8 44.2 3.4
e
2.45 0.99 0.83 4.28 4.53 3.29
NS NS NS NS P < 0.05 NS
Quadratic NS NS NS NS NS NS
Control (CON) = 0 g/kg physic nut meal (PNM), 100 g/kg PNM (PNM100), 200 g/kg PNM (PNM200), or 300 g/kg PNM (PNM300). SD, standard deviation. FBW, fasting body weight; HCW, hot carcass weight; CCW, cold carcass weight; HCY, hot carcass yield; CCY, cold carcass yield; CLI, cooling loss index. The means presented here were not significantly different (P > 0.05) among diets. NS, not significant (P > 0.05).
The FBW, HCW, CCW, HCY and CLI (Table 3) as well as carcass and meat cut weights did not differ (P > 0.05) between the diets (Table 4). Similarly, no significant differences (P > 0.05) were detected in the hemogram and serum AST, GGT, creatinine and albumin between diets as shown in Tables 5 and 6.
4. Discussion Mean dry matter intake observed in this study was similar to the NRC (2007) recommendation for the late maturing lambs: 0.83–1.20 kg/d for 20–30 kg BW animals. Therefore, non-toxic PNM was well accepted by the
Table 4 Measurements of carcass and the meat cuts weights of Santa Inês lambs fed diets with or without non-toxic physic nut meal. Dietsa
Half-carcass (kg) Legc (kg) Rib (kg) Shoulder (kg) Loin (kg) Neck (kg) Flank steak (kg) Skin weight (kg) Testicles (g) Scrotal circumference (cm) Thoracic viscera (kg) Abdominal viscera (kg) a b c d e
CON
PNM100
PNM200
PNM300
SDb
5.45d 1.82 1.44 1.06 0.37 0.47 0.29 2.08 246.67 21.8 0.94 0.58
5.53 1.82 1.41 1.09 0.38 0.50 0.32 1.98 250.00 21.5 0.97 0.60
5.29 1.80 1.38 1.02 0.34 0.45 0.29 2.10 211.70 20.9 1.00 0.60
5.15 1.74 1.32 1.00 0.30 0.50 0.28 2.21 243.33 23.0 0.90 0.55
0.48 0.04 0.05 0.01 0.02 0.02 0.01 0.26 10.11 1.56 0.02 0.02
Regression Linear
Quadratic
NSe NS NS NS NS NS NS NS NS NS NS NS
NS NS NS NS NS NS NS NS NS NS NS NS
Control (CON) = 0 g/kg physic nut meal (PNM), 100 g/kg PNM (PNM100), 200 g/kg PNM (PNM200), or 300 g/kg PNM (PNM300). SD, standard deviation. Meat cuts made in the left half-carcass. The means presented here were not significantly different (P > 0.05) among diets. NS, not significant (P > 0.05).
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Table 5 Hemogram of lambs fed diets with or without non-toxic physic nut meal. Hemogram
PCVc (%) Erythrocytes (×106 /L) Hemoglobin (g/dL) Leukocytes (×103 /mL) Platelets (×103 /mL) a b c d e
Dietsa CON
PNM100
PNM200
PNM300
SDb
Reference values
32.3d 9.8 10.3 7.3 587.8
31.2 9.4 10.2 9.4 657.7
31.0 9.5 9.8 9.5 593.7
31.2 9.6 9.8 10.2 612.8
3.49 1.08 3.02 2.63 96.48
27–45 9–15 9–15 4–12 205–705
Regression Linear
Quadratic
NSe NS NS NS NS
NS NS NS NS NS
Control (CON) = 0 g/kg physic nut meal (PNM), 100 g/kg PNM (PNM100), 200 g/kg PNM (PNM200), or 300 g/kg PNM (PNM300). SD, standard deviation. PCV, packed cell volume. The means presented here were not significantly different (P > 0.05) among diets. NS, not significant (P > 0.05).
Table 6 Serum variables of lambs fed diets with or without non-toxic physic nut meal. Serum variables
AST (U/L)c GGT (U/L) Creatinine (mg/100 dL) Albumin (g/100 mL) a b c d e
Dietsa CON
PNM100
PNM200
PNM300
SDb
135.0d 51.5 1.3 3.3
110.7 49.7 1.3 2.8
100.2 34.3 1.3 3.2
129.8 44.2 1.3 2.4
31.24 19.53 0.18 1.82
Reference values
98–278 20–52 1.2–1.9 2.4–3.9
Regression Linear
Quadratic
NSe NS NS NS
NS NS NS NS
Control (CON) = 0 g/kg physic nut meal (PNM), 100 g/kg PNM (PNM100), 200 g/kg PNM (PNM200), or 300 g/kg PNM (PNM300). SD, standard deviation. AST, aspartate aminotransferase; GGT, gamma-glutamyl transferase. The means presented here were not significantly different (P > 0.05) among diets. NS, not significant (P > 0.05).
lambs since no significant statistical difference in DMI was detected between the CON and the other diets. On the other hand, Araújo et al. (2010) using 0, 150, 300 and 450 g/kg of physic nut hull in the diet of cross bred sheep, observed a reduction in DMI as the level of inclusion of physic nut in the diet was increased, however, in support of this observation, Makkar et al. (1998) reported that the low physic nut acceptability was due to the presence of high concentrations of phorbol ester. The ADG (139.3, 123.8, 126.2 and 123.8 g/day/animal for CON, PNM100, PNM200 and PNM300, respectively) were similar to those found by Louvandini et al. (2007) – 139 g/day/animal, and Sousa et al. (2008) – 107 g/day/animal, feeding Santa Inês lambs with concentrate mixtures based on ground corn and soybean meal; and ground corn, soybean meal and wheat meal, respectively. Also, there was no difference in feed conversion ratio among diets, suggesting that this feed may be useful for sheep feeding and confirming that there were no acceptability problems for the non-toxic PNM. Although animals presented similar FBW, CCY showed a positive linear regression with increasing the level of PNM. Cold carcass yield was the only variable that presented this behavior; therefore more research is needed for a better understanding of this phenomenon. Sousa et al. (2009) obtained 45.3, 42.4% for HCY and CCY respectively, in Santa Inês lambs fed a 40% forage and 60% concentrate diet, and an average body weight of 30 kg at the slaughter. However, in the current, lambs had HCY and CCY values varying from 39.9 to 45.8% and 38.3 to 44.2%, respectively, which
is considered normal as explained by Pinheiro et al. (2009) who observed that carcass yield is affected by a number of inherent and non-inherent animal factors, such as slaughter weight and animal breed. Other authors have found higher HCY values, such as Garcia et al. (2000) who obtained a HCY of 53% in feedlot Santa Inês lambs fed coffee hull. Urano et al. (2006), found, for Santa Inês lambs fed different levels of soybean grain in the diet, an average for HCY and CCY of 49 and 48%, respectively. The yield levels found in the present study are slightly lower than that reported by the literature. This may be explained by the fact that the final body weight influences yield (Furusho-Garcia et al., 2004). Other trials had animals slaughtered with body weight above 30 kg while in this study, the animals were slaughtered with mean final body weight of 28.7 kg. Genetic studies (McManus et al., 2010) have shown that Brazilian Santa Inês sheep can be divided into two distinct sub-groups related to recent crossbreeding. One group has been crossed with terminal sire breeds to improve carcass quality, while the other is a relatively unimproved group. This may also explain the low carcass yield found here. Trials with ruminants fed physic nut are commonly not carried out long enough to evaluate growth performance and carcass characteristics, since animals show clinical signs of intoxication and some deaths occur at early stages of the experiments (Ahmed and Adam, 1979; Abdel Gadir ´ et al., 2010). et al., 2003; Araujo The cooling loss index, which represents the weight loss during carcass refrigeration (Galvani et al., 2008), is mainly
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influenced by the carcass fat cover and moisture loss in the cold chamber (Cunha et al., 2008). As there was no difference between the CLI of the groups which received PNM in the diet and CON, it may be inferred that the levels of PNM used in the trial had no influence on the carcass fat cover of the animals. Carcass and cuts weights showed no significant difference between groups, demonstrating that the treatments with different levels of PNM had the same meat production as the control group. The values obtained were similar to those obtained by other authors that studied carcass characteristics of feedlot Santa Inês sheep, such as Cunha et al. (2008) and Louvandini et al. (2006). Comparing the four treatments used in this experiment, it was seen that the animals fed a diet prepared with soybean meal (control group) had the same performance, carcass traits and meat cuts weights as the animals fed up to 300 g/kg inclusion of non-toxic physic nut meal in the concentrate. Therefore it is suggested that the residue of non-toxic physic nut (with zero concentration of phorbol ester), previously discarded by the industry without any economic value, may be employed in the nutrition of sheep and satisfactory animal performance can be achieved. The results of the blood tests conducted in this experiment showed no difference between the treatments, and all the results were within the reference range established by Pugh (2005). Therefore, there was no treatment effect for the variables: number of erythrocytes, hemoglobin concentration, leukocyte count, PCV and the number of platelets per mL of blood. Serum concentrations of AST, GGT, creatinine and albumin were not different between the diets. All evaluations performed presented results within reference range (Viana, 2007). These results showed that there was no liver or kidney damage through inclusion of this product up to 300 g/kg DM in concentrate mixture. Abdel Gadir et al. (2003), evaluating the toxicity of physic nut in Nubian goats kids, reported an increase in AST levels and decrease in packed cell volume, erythrocytes, hemoglobin and leukocytes. In addition, clinical symptoms of intoxication by physic nut were observed after three days on experiment. These included diarrhea, dyspnea, dehydration, in appetence and weakness. Clinical symptoms of intoxication and alterations in blood and serum biochemical variables may be attributed to the fact that these authors probably worked with a toxic variety of physic nut, with high concentrations of phorbol ester in its composition. This compound was not detected in the PNM used in the present study, so alterations in blood and serum biochemical variables as well as clinical symptoms of intoxication were not observed throughout the whole experimental period.
5. Conclusions The non-toxic physic nut meal is a promising alternative feedstuff for sheep nutrition. It may be employed in diet at inclusion levels of up to 300 g/kg DM in concentrate, causing no harmful effects on lamb performance, carcass characteristics or intoxication symptoms.
Acknowledgements The author thanks Informac¸ão Genético-Sanitária da Pecuária Brasileira/INCT/CNPq (National Counsel of Technological and Scientific Development) and CAPES (Coordination for the Improvement of Higher Level Personnel) for financial support. This experiment has been approved by the ethics committee of the University of Brasília, protocol number 33/2009.
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Sousa, V.S., Louvandini, H., Scropfner, E.S., Mcmanus, C.M., Abdalla, A.L., Garcia, J.A.S., 2008. Desempenho, características de carcac¸a e componentes corporais de ovinos deslanados alimentados com silagem de girassol e silagem de milho. Cienc. Anim. Bras. 9, 284–291. Sousa, W.H., Brito, E.A., Medeiros, A.N., Cartaxo, F.Q., Cezar, M.F., Cunha, M.G.G.C., 2009. Características morfométricas e de carcac¸a de cabritos e cordeiros terminados em confinamento. Rev. Bras. Zootec. 38, 1340–1346. Souza, A.D.V., Fávaro, S.P., Ítavo, L.C.V., Roscoe, R., 2009. Caracterizac¸ão química de sementes e tortas de pinhão-manso, nabo-forrageiro e crambe. Pesqui. Agropecu. Bras. 44, 1328–1335. Urano, F.S., Pires, A.V., Susin, I., Mendes, C.Q., Araujo, R.C., Mattos, W.R.S., 2006. Desempenho e características da carcac¸a de cordeiros confinados alimentados com grão de soja. Pesqui. Agropecu. Bras. 41, 1525–1530. Viana, F.A.B., 2007. Guia terapêutico veterinário, second ed. Cem, Lagoa Santa.
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Small Ruminant Research journal homepage: www.elsevier.com/locate/smallrumres
Growth and carcass characteristics of Santa Inês lambs fed diet supplemented with physic nut meal free of phorbol ester P.B. Oliveira a , P.M.T. Lima a,∗ , A. Campeche b , S. Mendonc¸a c , B.G. Laviola c , C. McManus a , H. Louvandini b a b c
Universidade de Brasília, Campus Universitário Darcy Ribeiro, 70910900 Brasília, DF, Brazil Centro de Energia Nuclear na Agricultura, Universidade de São Paulo, Av. Centenário, no 303, 13416000 Piracicaba, SP, Brazil Embrapa Agroenergia, Parque Estac¸ão Biológica, 70770901 Brasília, DF, Brazil
a r t i c l e
i n f o
Article history: Received 11 June 2012 Received in revised form 13 May 2013 Accepted 14 May 2013 Available online 10 June 2013 Keywords: Bio-fuel By-product Carcass Jatropha curcas
a b s t r a c t The aim of this work was to evaluate the growth performance, carcass traits, hematological and biochemical variables of Santa Inês lambs fed diet supplemented with physic nut (Jatropha curcas) meal (PNM). Twenty-four intact male lambs [age: 120 ± 0.98 days; body weight (BW): 19.7 ± 2.4 kg] were assigned randomly to four concentrate mixtures. These mixtures were supplemented with 0 (CON), 100 (PNM100), 200 (PNM200), or 300 (PNM300) g/kg dry matter (DM) of PNM. The concentrate was offered at 1.2% BW, whereas lambs were fed Tifton (Cyndon dactylon CV. Tifton-85) hay ad libitum. Animals were weighed and blood collected every 15 days. At the end of the experiment, lambs were slaughtered and their carcasses were evaluated. Biochemical and hematological variables were not different among diets (P > 0.05). Similarly, dry matter intake was not different among diets (P > 0.05) and no acceptability problems were observed for the PNM. Average daily gain and carcass characteristics also did not differ (P > 0.05) between the treatments. No clinical symptoms of intoxication by physic nut were detected during the whole experimental period. Therefore, it may be inferred that the non-toxic PNM was shown to be a promising protein source for sheep nutrition and can be used in concentrations of up to 300 g/kg DM in concentrate mixture. © 2013 Elsevier B.V. All rights reserved.
1. Introduction Agro-industry by-products have been used mainly to reduce production costs, and to offset increasing environmental concerns about less expensive waste management programs (Grasser et al., 1995). Studies have demonstrated that residues of the bio-fuel industry may have satisfactory levels of energy and protein, supplying requirements and
∗ Corresponding author at: Centro de Energia Nuclear na Agricultura, ´ no 303, 13416000, Piracicaba, Universidade de S˜ao Paulo, Av. Centenario, SP, Brazil. Tel.: +55 061 82784646; fax: +55 019 34294728. E-mail addresses: [email protected], [email protected] (P.M.T. Lima). 0921-4488/$ – see front matter © 2013 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.smallrumres.2013.05.007
substituting commonly used ingredients, such as soybean meal and corn grains. Among these residues, physic nut meal (PNM) is not widely used, as it is regarded as toxic for animals (Abdalla et al., 2008). It was previously believed that the toxicity of the physic nut is due to the presence of curcin, which has a toxic activity (Ribosome-Inactivating Protein – RIP) similar to the castor bean (Ricinus communis) ricin (Felix et al., 2008; Mendonca and Laviola, 2009). However, recent research has shown that the toxicity of physic nut seeds and oil is mainly due to the presence of phorbol ester, and not to curcin (Abdalla et al., 2008). One of the great advantages of physic nut is its production cycle, which may last for 40 years, maintaining an average productivity of two tons per hectare (Mendonca
P.B. Oliveira et al. / Small Ruminant Research 114 (2013) 20–25
and Laviola, 2009). The physic nut seed has approximately 210 g/kg crude protein (CP) and 400 g/kg ether extract (EE), while PNM has 286 g/kg CP and 142 g/kg EE on dry matter basis (Souza et al., 2009). Presently, there are more than 30 thousand hectares of physic nut in Brazil, distributed over the Southeast, Center-West and Northeast regions of the country. Considering only the plants that have reached the mature stage, there is a potential for producing more than 90 thousand tons of seeds per year, which would correspond to 58.5 thousand tons of PNM per year (Mendonca and Laviola, 2009). With new detoxification techniques and varieties of physic nut with zero concentration of phorbol ester (nontoxic), new uses for these residues arise. This protein rich feed (detoxified or non-toxic), which has phorbol ester as its limiting factor, is still not well studied for use as a feedstuff in animal nutrition. Generally, pressedcakes or meals generated by the oil extraction process do not pass through any kind of value-adding procedure, and their economic and nutritional potential, except for some materials from soybean or cottonseed processing, are unknown (Abdalla et al., 2008; Mendonca and Laviola, 2009). The objective of this study was to evaluate growth performance, carcass traits, as well as hematological and serum biochemical variables of Santa Inês lambs fed different levels of non-toxic physic nut meal in the diet. 2. Materials and methods 2.1. Study area The experiment was carried out at the Sheep Management Center, in the Água Limpa Experimental Station of the University of Brasília. The animals were allocated in individual pens of 1.8 m2 each, disposed in a 240 m2 shed.
2.2. Animals and diets Twenty-four male Santa Inês lambs [age: 120 ± 0.98 days; body weight (BW): 19.7 ± 2.4 kg] were divided into four diets (6 lambs/diet). Diets were: (1) control (270 g/kg soybean meal and 730 g/kg ground corn; CON); (2) 220 g/kg soybean meal, 680 g/kg ground corn and 100 g/kg PNM; PNM100; (3) 180 g/kg soybean meal, 620 g/kg ground corn and 200 g/kg PNM; PNM200; and (4) 140 g/kg soybean meal, 560 g/kg ground corn, and 300 g/kg PNM; PNM300. The experimental period lasted for 70 days. The physic nut meal (PNM) that was used in the current study was considered not toxic because phorbol ester was not detected when analyzed according to procedure described by Makkar and Becker (1999). The PNM (Germplasm Bank of Embrapa Agroenergy – CNPAE-169 and CNPAE-170) was purchased from a rural property in Mato Grosso do Sul State and was provided for the experiment by Embrapa Agroenergy after oil extraction by mechanical press. Each lamb was allocated to an individual pen (1.60 m × 1.15 m). The diets were balanced to offer 180 g/kg of CP and 800 g/kg of total digestive nutrients. The PNM was included in each diet as a substitute of total protein. Lambs had ad libitum access to water and Tifton (Cyndon dactylon CV. Tifton-85) hay. The chemical composition of the hay was 872.0, 68.8, 759.0, 410.0, 7.9, 51.0, and 534 g/kg DM, CP, neutral detergent fiber (NDF), acid detergent fiber (ADF), ether extract (EE), ash and total digestible nutrients (TDN), respectively. The concentrate was offered at 1.2% BW. The forage and concentrate mixture were offered separately, so that the PNM intake could be precisely calculated. The chemical analyses of the Tifton hay and concentrates were performed according to AOAC (1995) and Mertens (2002).
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Table 1 Chemical composition of concentrates used in the diets. Nutrient (g/kg)
Dry matter Crude protein Neutral detergent fiber Acid detergent fiber Ether extract Ash Total digestible nutrientsb
Dietsa CON
PNM100
PNM200
PNM300
879.0 201.0 108.0 57.0 35.1 19.9 810.0
883.0 200.8 140.0 70.2 38.9 18.7 808.0
887.0 200.5 171.4 88.4 42.1 17.9 807.0
890.0 200.3 200.1 106.0 45.5 17.2 807.0
a Control (CON) = 0 g/kg physic nut meal (PNM), 100 g/kg PNM (PNM100), 200 g/kg PNM (PNM200), and 300 g/kg PNM (PNM300). b Calculated according to Harlan et al. (1991).
To calculate dry matter intake (DMI), the leftovers of forage were collected and weighed three times a week. There were no concentrate leftovers. Lambs were weighed fortnightly, before they received the concentrate in the morning, throughout experimental period. After weighing, blood was collected via jugular venipuncture using vacuum tubes with and without anticoagulant (EDTA). The hemogram analyses were performed using an ABX – model 22P ABX micro 60® automatic counter. Biochemical tests (aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), creatinine and albumin) were performed using blood serum and analyses were carried out using commercial biochemical tests kits Labtest® and spectrophotometer Bioplus, model Bio-2000® . On the last day of the experimental period, the animals were slaughtered so that the carcass characteristics could be evaluated, according to procedures described by Osório et al. (1998). The slaughtering took place after a 16 h fast, following the current regulations in Brazil, Regulation of Industrial and Sanitary Inspection for Products of Animal Origin – RIISPOA (1952). Immediately before the slaughtering, animals were weighed to obtain fasting body weight (FBW). The animals were stunned by electrocution. After bleeding, skinning and evisceration, the viscera were weighed in two groups: Group A (rumen, reticulum, omasum, abomasum and intestines) and Group B (tongue, esophagus, trachea, lungs, heart, liver and kidneys). During slaughter, scrotal circumference was also measured. The whole animal carcass was used to determine hot carcass weight (HCW), and hot carcass yield (HCY), using the equation: HCY = HCW/FBW × 100. Carcasses were cooled in cold chamber, at temperatures below 4 ◦ C + 2 ◦ C for 24 h and weighed again to obtain cold carcass weight (CCW). Cold carcass yield (CCY) was calculated: CCY = CCW/FBW × 100 and the cooling loss index, CLI = (HCW − CCW)/HCW × 100. A longitudinal cut was made along the vertebral column, splitting the carcass in two parts, which were then weighed. The carcass was cut into the following parts: neck, shoulder, loin, ribs, flank steak, and leg which were weighed individually. 2.3. Statistical analysis The experiment was conducted in a completely randomized design. The data analysis was carried out by the software SAS v.9.2® (Cary, North Carolina), using the MIXED procedure for measures repeated over time. For the variables measured only once, the GLM procedure was employed using ANOVA analysis. The REG procedure was used to study the effects of increasing PMN in the diet over all the variables evaluated. For all the analysis, the significance level adopted was 5% (P < 0.05).
3. Results The chemical composition of concentrates used in the diet is presented in Table 1. Growth performance of Santa Inês lambs fed diets with or without physic nut meal is shown in Table 2. No significant differences (P > 0.05) were observed in DMI, average daily gain (ADG), and feed conversion ratio between diets.
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Table 2 Growth performance of Santa Inês lambs fed diets containing physic nut meal. Dietsa
Variables
Duration (days) Initial BWc (kg) Final BW (kg) ADG (g/animal/day) DMI (g/animal/day) FCR (g gain/kg feed) a b c d e
CON
PNM100
PNM200
PNM300
SDb
70 19.6d 29.4 139.3 1053.8 132.2
70 19.5 28.2 123.8 1002.1 126.4
70 19.8 28.6 126.2 1013.8 124.5
70 19.8 28.5 123.8 1010.7 122.5
– 2.4 2.5 22.2 214.2 21.17
Regression Linear
Quadratic
– NSe NS NS NS NS
– NS NS NS NS NS
Control (CON) = 0 g/kg physic nut meal (PNM), 100 g/kg PNM (PNM100), 200 g/kg PNM (PNM200), or 300 g/kg PNM (PNM300). SD, standard deviation. BW, body weight; ADG, average daily gain; DMI, dry matter intake; FCR, feed conversion ratio. The means presented here were not significantly different (P > 0.05) among diets. NS, not significant (P > 0.05).
Table 3 Carcass traits of Santa Inês lambs fed diets with or without physic nut meal. Dietsa CON
PNM100
PNM200
PNM300
SDb
Regression Linear
c
FBW (kg) HCW (kg) CCW (kg) HCY (%) CCY (%) CLI (%) a b c d e
d
26.7 10.6 10.2 39.9 38.3 3.7
26.6 11.0 10.6 41.7 40.3 3.2
27.3 11.2 10.8 41.0 39.6 3.0
25.2 11.5 11.1 45.8 44.2 3.4
e
2.45 0.99 0.83 4.28 4.53 3.29
NS NS NS NS P < 0.05 NS
Quadratic NS NS NS NS NS NS
Control (CON) = 0 g/kg physic nut meal (PNM), 100 g/kg PNM (PNM100), 200 g/kg PNM (PNM200), or 300 g/kg PNM (PNM300). SD, standard deviation. FBW, fasting body weight; HCW, hot carcass weight; CCW, cold carcass weight; HCY, hot carcass yield; CCY, cold carcass yield; CLI, cooling loss index. The means presented here were not significantly different (P > 0.05) among diets. NS, not significant (P > 0.05).
The FBW, HCW, CCW, HCY and CLI (Table 3) as well as carcass and meat cut weights did not differ (P > 0.05) between the diets (Table 4). Similarly, no significant differences (P > 0.05) were detected in the hemogram and serum AST, GGT, creatinine and albumin between diets as shown in Tables 5 and 6.
4. Discussion Mean dry matter intake observed in this study was similar to the NRC (2007) recommendation for the late maturing lambs: 0.83–1.20 kg/d for 20–30 kg BW animals. Therefore, non-toxic PNM was well accepted by the
Table 4 Measurements of carcass and the meat cuts weights of Santa Inês lambs fed diets with or without non-toxic physic nut meal. Dietsa
Half-carcass (kg) Legc (kg) Rib (kg) Shoulder (kg) Loin (kg) Neck (kg) Flank steak (kg) Skin weight (kg) Testicles (g) Scrotal circumference (cm) Thoracic viscera (kg) Abdominal viscera (kg) a b c d e
CON
PNM100
PNM200
PNM300
SDb
5.45d 1.82 1.44 1.06 0.37 0.47 0.29 2.08 246.67 21.8 0.94 0.58
5.53 1.82 1.41 1.09 0.38 0.50 0.32 1.98 250.00 21.5 0.97 0.60
5.29 1.80 1.38 1.02 0.34 0.45 0.29 2.10 211.70 20.9 1.00 0.60
5.15 1.74 1.32 1.00 0.30 0.50 0.28 2.21 243.33 23.0 0.90 0.55
0.48 0.04 0.05 0.01 0.02 0.02 0.01 0.26 10.11 1.56 0.02 0.02
Regression Linear
Quadratic
NSe NS NS NS NS NS NS NS NS NS NS NS
NS NS NS NS NS NS NS NS NS NS NS NS
Control (CON) = 0 g/kg physic nut meal (PNM), 100 g/kg PNM (PNM100), 200 g/kg PNM (PNM200), or 300 g/kg PNM (PNM300). SD, standard deviation. Meat cuts made in the left half-carcass. The means presented here were not significantly different (P > 0.05) among diets. NS, not significant (P > 0.05).
P.B. Oliveira et al. / Small Ruminant Research 114 (2013) 20–25
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Table 5 Hemogram of lambs fed diets with or without non-toxic physic nut meal. Hemogram
PCVc (%) Erythrocytes (×106 /L) Hemoglobin (g/dL) Leukocytes (×103 /mL) Platelets (×103 /mL) a b c d e
Dietsa CON
PNM100
PNM200
PNM300
SDb
Reference values
32.3d 9.8 10.3 7.3 587.8
31.2 9.4 10.2 9.4 657.7
31.0 9.5 9.8 9.5 593.7
31.2 9.6 9.8 10.2 612.8
3.49 1.08 3.02 2.63 96.48
27–45 9–15 9–15 4–12 205–705
Regression Linear
Quadratic
NSe NS NS NS NS
NS NS NS NS NS
Control (CON) = 0 g/kg physic nut meal (PNM), 100 g/kg PNM (PNM100), 200 g/kg PNM (PNM200), or 300 g/kg PNM (PNM300). SD, standard deviation. PCV, packed cell volume. The means presented here were not significantly different (P > 0.05) among diets. NS, not significant (P > 0.05).
Table 6 Serum variables of lambs fed diets with or without non-toxic physic nut meal. Serum variables
AST (U/L)c GGT (U/L) Creatinine (mg/100 dL) Albumin (g/100 mL) a b c d e
Dietsa CON
PNM100
PNM200
PNM300
SDb
135.0d 51.5 1.3 3.3
110.7 49.7 1.3 2.8
100.2 34.3 1.3 3.2
129.8 44.2 1.3 2.4
31.24 19.53 0.18 1.82
Reference values
98–278 20–52 1.2–1.9 2.4–3.9
Regression Linear
Quadratic
NSe NS NS NS
NS NS NS NS
Control (CON) = 0 g/kg physic nut meal (PNM), 100 g/kg PNM (PNM100), 200 g/kg PNM (PNM200), or 300 g/kg PNM (PNM300). SD, standard deviation. AST, aspartate aminotransferase; GGT, gamma-glutamyl transferase. The means presented here were not significantly different (P > 0.05) among diets. NS, not significant (P > 0.05).
lambs since no significant statistical difference in DMI was detected between the CON and the other diets. On the other hand, Araújo et al. (2010) using 0, 150, 300 and 450 g/kg of physic nut hull in the diet of cross bred sheep, observed a reduction in DMI as the level of inclusion of physic nut in the diet was increased, however, in support of this observation, Makkar et al. (1998) reported that the low physic nut acceptability was due to the presence of high concentrations of phorbol ester. The ADG (139.3, 123.8, 126.2 and 123.8 g/day/animal for CON, PNM100, PNM200 and PNM300, respectively) were similar to those found by Louvandini et al. (2007) – 139 g/day/animal, and Sousa et al. (2008) – 107 g/day/animal, feeding Santa Inês lambs with concentrate mixtures based on ground corn and soybean meal; and ground corn, soybean meal and wheat meal, respectively. Also, there was no difference in feed conversion ratio among diets, suggesting that this feed may be useful for sheep feeding and confirming that there were no acceptability problems for the non-toxic PNM. Although animals presented similar FBW, CCY showed a positive linear regression with increasing the level of PNM. Cold carcass yield was the only variable that presented this behavior; therefore more research is needed for a better understanding of this phenomenon. Sousa et al. (2009) obtained 45.3, 42.4% for HCY and CCY respectively, in Santa Inês lambs fed a 40% forage and 60% concentrate diet, and an average body weight of 30 kg at the slaughter. However, in the current, lambs had HCY and CCY values varying from 39.9 to 45.8% and 38.3 to 44.2%, respectively, which
is considered normal as explained by Pinheiro et al. (2009) who observed that carcass yield is affected by a number of inherent and non-inherent animal factors, such as slaughter weight and animal breed. Other authors have found higher HCY values, such as Garcia et al. (2000) who obtained a HCY of 53% in feedlot Santa Inês lambs fed coffee hull. Urano et al. (2006), found, for Santa Inês lambs fed different levels of soybean grain in the diet, an average for HCY and CCY of 49 and 48%, respectively. The yield levels found in the present study are slightly lower than that reported by the literature. This may be explained by the fact that the final body weight influences yield (Furusho-Garcia et al., 2004). Other trials had animals slaughtered with body weight above 30 kg while in this study, the animals were slaughtered with mean final body weight of 28.7 kg. Genetic studies (McManus et al., 2010) have shown that Brazilian Santa Inês sheep can be divided into two distinct sub-groups related to recent crossbreeding. One group has been crossed with terminal sire breeds to improve carcass quality, while the other is a relatively unimproved group. This may also explain the low carcass yield found here. Trials with ruminants fed physic nut are commonly not carried out long enough to evaluate growth performance and carcass characteristics, since animals show clinical signs of intoxication and some deaths occur at early stages of the experiments (Ahmed and Adam, 1979; Abdel Gadir ´ et al., 2010). et al., 2003; Araujo The cooling loss index, which represents the weight loss during carcass refrigeration (Galvani et al., 2008), is mainly
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influenced by the carcass fat cover and moisture loss in the cold chamber (Cunha et al., 2008). As there was no difference between the CLI of the groups which received PNM in the diet and CON, it may be inferred that the levels of PNM used in the trial had no influence on the carcass fat cover of the animals. Carcass and cuts weights showed no significant difference between groups, demonstrating that the treatments with different levels of PNM had the same meat production as the control group. The values obtained were similar to those obtained by other authors that studied carcass characteristics of feedlot Santa Inês sheep, such as Cunha et al. (2008) and Louvandini et al. (2006). Comparing the four treatments used in this experiment, it was seen that the animals fed a diet prepared with soybean meal (control group) had the same performance, carcass traits and meat cuts weights as the animals fed up to 300 g/kg inclusion of non-toxic physic nut meal in the concentrate. Therefore it is suggested that the residue of non-toxic physic nut (with zero concentration of phorbol ester), previously discarded by the industry without any economic value, may be employed in the nutrition of sheep and satisfactory animal performance can be achieved. The results of the blood tests conducted in this experiment showed no difference between the treatments, and all the results were within the reference range established by Pugh (2005). Therefore, there was no treatment effect for the variables: number of erythrocytes, hemoglobin concentration, leukocyte count, PCV and the number of platelets per mL of blood. Serum concentrations of AST, GGT, creatinine and albumin were not different between the diets. All evaluations performed presented results within reference range (Viana, 2007). These results showed that there was no liver or kidney damage through inclusion of this product up to 300 g/kg DM in concentrate mixture. Abdel Gadir et al. (2003), evaluating the toxicity of physic nut in Nubian goats kids, reported an increase in AST levels and decrease in packed cell volume, erythrocytes, hemoglobin and leukocytes. In addition, clinical symptoms of intoxication by physic nut were observed after three days on experiment. These included diarrhea, dyspnea, dehydration, in appetence and weakness. Clinical symptoms of intoxication and alterations in blood and serum biochemical variables may be attributed to the fact that these authors probably worked with a toxic variety of physic nut, with high concentrations of phorbol ester in its composition. This compound was not detected in the PNM used in the present study, so alterations in blood and serum biochemical variables as well as clinical symptoms of intoxication were not observed throughout the whole experimental period.
5. Conclusions The non-toxic physic nut meal is a promising alternative feedstuff for sheep nutrition. It may be employed in diet at inclusion levels of up to 300 g/kg DM in concentrate, causing no harmful effects on lamb performance, carcass characteristics or intoxication symptoms.
Acknowledgements The author thanks Informac¸ão Genético-Sanitária da Pecuária Brasileira/INCT/CNPq (National Counsel of Technological and Scientific Development) and CAPES (Coordination for the Improvement of Higher Level Personnel) for financial support. This experiment has been approved by the ethics committee of the University of Brasília, protocol number 33/2009.
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