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Effect of substitution of soybean meal with treated or untreated high glucosinolate mustard (Brassica juncea) meal on intake, digestibility, growth performance and body composition of calves
Animal Feed Science and Technology 94 (2001) 137±146
Effect of substitution of soybean meal with treated or untreated high glucosinolate mustard (Brassica juncea) meal on intake, digestibility, growth performance and body composition of calves M.K. Tripathia,*, I.S. Agrawalb, S.D. Sharmab, D.P. Mishrac a
Division of Animal Nutrition, Central Sheep and Wool Research Institute, Avikanagar, Rajasthan 304501, India b Department of Animal Science, Govind Ballabh Pant University of Agriculture and Technology, Pantnagar 263145, UP, India c Department of Biochemistry, Govind Ballabh Pant University of Agriculture and Technology, Pantnagar 263145, UP, India Received 4 July 2000; received in revised form 14 August 2001; accepted 14 August 2001
Abstract Effects of feeding acid treated mustard (Brassica juncea) meal and untreated mustard meal (MM) as a source of protein on dry matter (DM) intake, digestibility, growth rate, body composition and blood constituents was studied in (Jersey Sahiwal) crossbred growing calves. Diets consisted of oat hay offered ad libitum with a compound feed mixture (CFM) as a supplement to provide protein and other nutrients. CFMs were formulated using soybean meal (SBM), untreated MM or acid treated MM. Differences among treatment groups in DM intake of oat hay and in total DM intake as a percentage of body weight (BW) favoured (P < 0:05) the SBM diet, whereas acid treated MM and untreated MM fed calves had similar DM intake. The feed conversion ratio was the lowest (P < 0:05), and growth rate was highest (P < 0:05), in calves fed SBM and acid treated MM diets. Serum albumin was lowest (P < 0:05) in calves fed the untreated MM diet. Acid treated MM can replace SBM as a protein source without substantive detrimental effects on overall calf performance and has bene®cial effects on performance of growing calves compared to untreated MM. # 2001 Published by Elsevier Science B.V. Keywords: Mustard meal; Glucosinolate; Growth; Body composition; Calves
* Corresponding author. Tel.: 91-1437-20143; fax: 91-1437-20163. E-mail address: [email protected] (M.K. Tripathi).
0377-8401/01/$ ± see front matter # 2001 Published by Elsevier Science B.V. PII: S 0 3 7 7 - 8 4 0 1 ( 0 1 ) 0 0 2 8 6 - 3
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1. Introduction Mustard (Brassica juncea) is the second most important group of oil seeds and India is the second largest producer of rapeseed mustard in the world (Kiresur, 1999), producing 20% of the world's rapeseed mustard. Mustard group oil seed crops have the advantage of being more draught tolerant and disease resistant than other oil seed crops and, with the recent increase in renovated dryland agriculture in India, the production of mustard has been increased substantially. Mustard meal (MM) is the by-product that remains following oil extraction from mustard seeds. On a dry matter (DM) basis (kg 1), MM has 459 g crude protein (CP), 212 g neutral detergent ®ber (NDF) and 128 g acid detergent ®ber (ADF: Newkirk et al., 1997). In spite of its high CP content and well balanced amino acid composition, the use of MM in animal feeding is limited because the meal contains glucosinolate (GLS), compounds that reduce nutritive value and makes MM unpalatable as well as goitrogenic. The GLS degradation products thiocyanate, isothiocyanate and nitriles suppress thyroid uptake of iodine and result in low levels of the thyroid hormones tri-iodothyronine and thyroxin (Barrett et al., 1997). The inclusion of GLS containing meal at higher levels induces metabolic disorders, such as liver and thyroid hypertrophy (Papas et al., 1979; Bell, 1984). These problems are associated with GLS degradation into toxic compounds either by myrosinase inherently present in the cellular compartment of feed or the enzyme present in bacterial micro¯ora (Nugon-Baudon et al., 1990). Most varieties of mustard grown in India contain 12±90 mg g 1 GLS meal (Chauhan et al., 1999). In order to remove or detoxify GLS and improve the nutritional value of rapeseed and mustard meals, acid treatment in combination with heat has been shown to destroy nearly 95% of intact GLS in MM (Tripathi and Agrawal, 1998). Mitigating the deleterious effects of GLS, allow MM to be used in livestock feeding. Our objective was to study the effect of replacing the soybean meal (SBM) with acid treated MM (AMM) and untreated MM (UMM) as protein sources on DM intake, nutrient utilisation, blood biochemical constituents, growth performance, and body composition of calves. 2. Materials and methods 2.1. Acid treatment of mustard meal Untreated MM contained 898.0 g DM, 282.8 g CP, 107.5 g kg 1 ether extract (EE) and 46.2 mg GLS g 1 oil extracted meal. The UMM was treated (Tripathi and Agrawal, 1998) with 16 ml HCl per kg, which raised its moisture content to 40%, after which it was allowed to stand for 72 h at room temperature followed by heating in a forced air oven at 1808C for 2 h. 2.2. Animals and diets Twenty four male crossbred (Jersey Sahiwal) growing calves of 240 15:4 days of age and 87:3 2:52 kg BW were dewormed with `Albendazole'(at 10 mg kg 1 BW) and
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divided into three equal groups based on comparable age and BW. The diet consisted of oat hay (OH) offered ad libitum and a CFM as a supplement to provide adequate protein and other nutrients (ICAR, 1998). The CFM was formulated using SBM or untreated MM or acid treated MM. Untreated MM and acid treated MM completely replaced SBM on a protein basis. Calves were housed individually in stalls and OH and CFM were offered separately. The calves were permitted 10% OH refusals. The daily allowance of CFM was calculated based on previous 14 days OH intake, rate of live weight gain and BW. 2.2.1. Growth trial The growth trial lasted for 16 weeks in a randomised design during which feed intake was recorded daily. Samples of OH and CFM offered were collected once weekly for estimation of DM, and 3 or 4 weeks samples were pooled for chemical analysis. The BW of the calves were recorded for three consecutive days at 14 days intervals before offering feed and water, and mean weights were used to determine growth. 2.2.2. Metabolism trial A metabolism trial was conducted after 40 days of experimental feeding using six calves in each treatment. The metabolism trial lasted for 7 days during which daily intake of OH and CFM, and output of faeces were recorded. Samples of feed offered, feed refusals, and faeces voided were collected every morning. The faeces excreted were collected using total collection bags. Two percent by weight of total faeces daily voided by individual animals was used for DM determination. DM in OH, CFM and faeces were determined by drying at 708C to a constant weight. The dried samples of each day for 7 days collection were pooled, ground through a 1 mm screen and preserved for chemical analysis. Samples of faeces (1/1000) from individual animals were collected every morning for 7 days in a 500 ml Kjeldahl ¯ask containing 25 ml concentrated sulphuric acid for N determination. 2.3. Blood sampling and analysis Blood samples were collected once at the end of the feeding period, through jugular puncture into heparinised tubes for haematological studies and into serum tubes for biochemical assays. Blood glucose, serum total proteins, and albumin were assayed by commercial kits (Span Diagnostics Ltd., New Delhi). Globulin was determined by subtracting albumin from total protein. The haematological measurements (haemoglobin, packed cell volume (PCV), erythrocyte sedimentation rate (ESR), white blood cell (WBC) and red blood cell count (RBC)) were completed by the procedures of Walmsley and White (1994). 2.4. Body composition At the end of the growth trial, animals were weighed for three consecutive days and the average weight was used to determine body composition. Body composition was determined using the antipyrine dilution technique (Brodie et al., 1949) and modi®ed by
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Wellington et al. (1956). Total body water was calculated (Soberman, 1950) from antipyrine level in plasma. The procedures of Bensadoum et al. (1963) for empty BW and Reid et al. (1963) for body fat and body protein estimation were used. The body mineral content was determined by subtracting the water and OM content of the body. 2.5. Chemical analysis Samples of OH, CFM and faeces were analysed for CP, EE and ash as per the procedures, numbered 988.05, 920.05 and 968.08, respectively, of AOAC (1995). The neutral detergent ®bre (NDF) was determined by the procedure of Van Soest et al. (1991) without sodium sulphite or a-amylase, whereas acid detergent ®bre (ADF) and acid detergent lignin (ADL) were determined according to the method described by Goering and Van Soest (1970). Metabolisable energy (ME) was estimated according to Kirchgessner (1995). Total GLS content in UMM, AMM and CFMs was determined using the thymol method (Tholen et al., 1989). Calcium was estimated by the titrimetric method, number 6.011 of AOAC (1965), and phosphorus by a colorimetric method (Donald et al., 1956). 2.6. Statistical analysis Data were analysed using analysis of variance technique (Snedecor and Cochran, 1968). Growth data were analysed by least square analysis (Harvey, 1975), with the model Yij m Ti eij where m is the general mean, Ti the effect of ith treatment (i 1, 3), eij is random error. 3. Results 3.1. Glucosinolate content of feeds The GLS content of UMM was 46.2 mg g 1 meal, which was reduced to 4.5 mg g 1 in AMM (Table 1). The GLS was 2.6 and 26.4 mg g 1 DM in CFM containing AMM and UMM, respectively. 3.2. Growth trial The BW gain was 56.6, 57.4 and 30.6 kg, respectively, in calves fed the diet with SBM, AMM and UMM and these differences were signi®cant (P < 0:05). Total BW gain and ADG was lower (P < 0:01) 26.0 kg and 220 g, respectively, in calves fed the diet with UMM compared with SBM or AMM calves. However, ADG, feed intake and FCR did not differ between SBM and AMM fed calves (Table 2). Feed intake was higher (P < 0:05) in SBM and AMM fed calves compared to UMM calves whereas FCR was lower in UMM fed calves. Calves fed the diet with UMM consumed 5.4±6.3 kg more (P < 0:01) feed for each kg gain to those fed the SBM or AMM diet, respectively.
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Table 1 Ingredient and chemical composition of compound feed mixtures (CFMs) of three treatments and hay used for experimental feeding Protein source in compound feed mixture Soybean meal
Oat hay
Mustard meal Acid treated
Untreated
1
Ingredient composition (g kg ) Soybean meal Acid treated mustard meal Untreated mustard meal Deoiled rice bran Wheat grain Salt Mineral mixturea
265 ± ± 350 355 10 20
± 495 ± 240 235 10 20
± ± 495 240 235 10 20
Chemical composition (g kg 1 of DM) Organic matter Crude protein Ether extract Neutral detergent fiber Acid detergent fiber Acid detergent lignin Calcium Phosphorus ME (MJ kg 1 DM)b Glucosinolate (mg g 1 DM)
860.5 216.3 15.2 509.0 167.0 35.5 17.0 12.4 11.06 NDc
870.3 207.5 16.2 499.0 166.0 47.0 20.0 11.6 11.48 2.6
871.8 216.0 16.9 481.0 168.0 50.2 22.0 11.5 11.78 26.4
883.5 147.5 20.0 609.5 388.5 57.0 16.5 1.6 9.02 NDc
a Composition: calcium 320 g kg 1, phosphorus 62 g kg 1, manganese 2.7 g kg 1, zinc 2.6 g kg 1, iron 1000 ppm, fluorine 900 ppm, iodine 100 ppm, copper 100 ppm. b Estimated according to Kirchgessner (1995). c Not detected.
Table 2 Growth performance of calves on the three treatmentsa Protein source in compound feed mixture Soybean meal
Initial body weight (kg) Final body weight (kg) Average daily gain (g per day) Total feed intake (kg) Feed conversion ratio (g/g) a
88.6 145.2 a 472 a 575 a 10.15 a
S.E.M.
Mustard meal Acid treated
Untreated
87.2 148.6 a 495 a 549 a 9.24 a
87.9 118.5 b 255 b 476 b 15.56 b
Values having different letters in rows differ significantly (P < 0:05).
5.3 6.8 57 21 1.04
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Table 3 Nutrient intake, digestibility and N utilisation of three diets used in metabolic triala Protein source in compound feed mixture Soybean meal
S.E.M.
Mustard meal Acid treated
Untreated
3.60 a 1.19 4.79 4.06 a 133.6 a
3.19 b 1.18 4.37 3.93 b 127.7 b
2.96 b 1.17 4.13 3.88 b 122.4 b
0.13 0.04 0.41 0.05 1.4
Metabolisable energy intake MJ per day
45.63
42.37
40.48
3.43
Digestibility (%) Dry matter Organic matter Crude protein Neutral detergent fiber Acid detergent fiber
63.1 65.1 65.3 61.3 50.7
61.8 63.7 66.9 58.6 45.6
62.5 64.4 67.6 57.8 49.8
1.6 1.2 1.3 1.4 2.0
Nitrogen utilisation Intake (g per day) Faecal (g per day) Absorbed (g per day) Absorbed (% intake)
128.5 a 45.3 83.2 a 64.7
121.9 b 40.6 73.9 b 64.5
108.5 b 34.1 74.4 b 68.6
2.0 4.4 2.2 1.9
Dry matter intake (kg per day) Hay (kg per day) Concentrateb (kg per day) Total (kg per day) (% BW) (g kg 1 W0.75)
a b
Values having different letters in rows differ significantly (P < 0:05). The relevant concentrate for the treatment (Table 1).
3.3. Metabolism trial Intake of DM in CFM, and in total, and ME intake were similar among the three groups (Table 3). The DMI of OH was higher (P < 0:01) in SBM fed calves compared to those fed AMM or UMM diets. 3.3.1. Nutrient digestibility and nitrogen utilisation Digestibility of DM constituents and ®ber fractions did not differ among diets. The N intake was lowest (P < 0:05) in calves fed the UMM diet compared to those fed SBM or AMM. The N voided in faeces showed the same trends as N intake. Absorption of N (g per day) was highest (P < 0:05) in calves fed SBM diet. 3.4. Blood biochemical constituents and body composition The albumin concentration was lowest in calves fed the UMM diet. Whereas, serum protein and blood glucose did not differ among diets. The ESR was highest in UMM, intermediate in AMM and lowest in SBM fed calves, Other haematological and body (i.e. water, protein, fat and mineral) constituents did not differ among treatment (Table 4).
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Table 4 Blood biochemical and haematological constituents, and body composition of calvesa Protein source in compound feed mixture Soybean meal
S.E.M.
Mustard meal Acid treated
Untreated
1
Blood constituents (mg dl ) Total serum protein Albumin Globulin Blood glucose
7.79 4.54 a 3.25 70
8.56 4.93 a 3.63 73
6.73 2.79 b 3.94 71
13.84 0.68 0.28 <1
Haematological constituents Haemoglobin (g%) Packed cell volume (%) Erythrocyte sedimentation rate (%) Red blood cell counts (106 mm 3) White blood cell counts (103 mm 3)
7.7 23.8 3.5 c 3.8 11.0
7.2 22.3 5.8 b 3.8 11.0
7.4 26.7 7.8 a 4.1 10.0
0.3 1.1 <0.1 0.2 0.5
Body composition (g kg Water Protein Fat Mineral matter a
1
empty body weight)
554 157 226 61
456 138 348 58
596 168 186 50
34 10 42 5
Values having different letters in rows differ significantly (P < 0:05).
4. Discussion Hydrolysis of GLS is caused by myrosinase enzyme, which is present in MM. Myrosinase expresses maximum activity at 15±40% moisture level (Mukherjee et al., 1976) and 3.5±8.0 pH (Gil and Macleod, 1980). In the present experiment, acid treatment of MM reduced GLS content sharply (90.3% GLS hydrolysed). The HCl treatment applied to MM increased the moisture level to 40% and changed the pH to favour catalytic activity of the myrosinase enzyme required for GLS degradation. Added HCl reduced the pH of the meal from 5.35 to 2.79 (Tripathi and Agrawal, 1998), which altered the nature of GLS degradation products. The heat applied during acid treatment (1808C for 2 h) also destroyed GLS and/or some of the GLS degradation products evaporated. Digestive processes in the rumen are the result of microbial and fungal activities, which are reduced due to the antifungal and antimicrobial activities of GLS and/or GLS degradation products (Manici et al., 1997). In contrast to this study, lower digestibility of DM constituents and ®ber fractions were reported in RSM (Sharma et al., 1980; Lardy, 1993) and MM (Tripathi et al., 2001a) diets. The metabolic trial in this study was conducted after only 40 days of experimental feeding. Possibly due to this short adaptation period, the known deleterious effects of GLS were less pronounced on the digestive ability of the calves, and so treatment differences failed to reach to statistical signi®cance. Tripathi et al. (2001b) reported that dietary glucosinolates exhibit deleterious effects on the digestive ability of calves, whereas glucosinolates require some time to manifest its
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deleterious effect on digestibility and growth. Lower DMI of calves fed untreated MM, compared to SBM or acid treated MM, was probably due to depressed appetite associated with the GLS or its degradation products (Pusztai, 1989; Duncan and Milne, 1990). Lower feed intake was also reported by Olsson (1978) and Tripathi et al. (1996, 1997) on RSM diets compared to SBM diets. Absorption of N containing compounds on acid treated MM diets might be higher in the small intestine because the heat applied during acid treatment of MM reduces degradability of protein in the rumen (érskov, 1981) thereby improving ef®ciency of amino acid absorption in the small intestine (Mustafa et al., 1999). The higher growth rate, better FCR and higher albumin concentration in calves fed the acid treated MM diet are probably the combined effects of higher N utilisation and negligible amounts of GLS. Lower albumin concentrations in untreated MM fed calves was probably the effect of reduced liver synthesis (Eggum, 1989; Walmsley and White, 1994; Rosenthal, 1997) due to the deleterious effects of GLS on liver function causing reduced N utilisation and poor growth. 5. Conclusion Mustard (B. juncea) meal, incorporated in diets of crossbred growing calves as a replacement for SBM negatively affected DM intake, growth rate, feed conversion ratio, N utilisation and serum albumin concentrations, presumably due to high the GLS content. However, when MM was treated with HCl, it could replace SBM without negative effects on N utilisation and growth performance. Acknowledgements The author wish to thank Director, Central Sheep and Wool Research Institute, Avikanagar for study leave and Dr. Mahender S. Rahal, Professor (Animal Nutrition), Department of Animal Science, Govind Ballabh Pant University of Agriculture and Technology, Pantnagar, for critical suggestions and cooperation during the experiment. References AOAC, 1965. Official Methods of Analysis, 10th Edition. Association of Official Analytical Chemists, Washington, DC. AOAC, 1995. Official Methods of Analysis, 16th Edition. Association of Official Analytical Chemists, Washington, DC. Barrett, J.E., Klopfenstein, C.F., Leipold, H.W., 1997. Detoxification of rapeseed meal by extrusion with an added basic salt. Cereal Chem. 74, 168±170. Bell, J.M., 1984. Nutrients and toxicants in rapeseed meal. A review. J. Anim. Sci. 4, 996±1010. Bensadoum, A., Van Niekerk, B.D.H., Paladines, D.L., Reid, J.T., 1963. Evaluation of antipyrine, N-acetyl-4aminoantipyrine and shrunk body weight in predicting the chemical composition of sheep body. J. Anim. Sci. 22, 604±612. Brodie, B.B., Axelrod, J., Soberman, R., Levy, B.B., 1949. The estimation of antipyrine in biological materials. J. Biol. Chem. 179, 25±29.
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Effect of substitution of soybean meal with treated or untreated high glucosinolate mustard (Brassica juncea) meal on intake, digestibility, growth performance and body composition of calves M.K. Tripathia,*, I.S. Agrawalb, S.D. Sharmab, D.P. Mishrac a
Division of Animal Nutrition, Central Sheep and Wool Research Institute, Avikanagar, Rajasthan 304501, India b Department of Animal Science, Govind Ballabh Pant University of Agriculture and Technology, Pantnagar 263145, UP, India c Department of Biochemistry, Govind Ballabh Pant University of Agriculture and Technology, Pantnagar 263145, UP, India Received 4 July 2000; received in revised form 14 August 2001; accepted 14 August 2001
Abstract Effects of feeding acid treated mustard (Brassica juncea) meal and untreated mustard meal (MM) as a source of protein on dry matter (DM) intake, digestibility, growth rate, body composition and blood constituents was studied in (Jersey Sahiwal) crossbred growing calves. Diets consisted of oat hay offered ad libitum with a compound feed mixture (CFM) as a supplement to provide protein and other nutrients. CFMs were formulated using soybean meal (SBM), untreated MM or acid treated MM. Differences among treatment groups in DM intake of oat hay and in total DM intake as a percentage of body weight (BW) favoured (P < 0:05) the SBM diet, whereas acid treated MM and untreated MM fed calves had similar DM intake. The feed conversion ratio was the lowest (P < 0:05), and growth rate was highest (P < 0:05), in calves fed SBM and acid treated MM diets. Serum albumin was lowest (P < 0:05) in calves fed the untreated MM diet. Acid treated MM can replace SBM as a protein source without substantive detrimental effects on overall calf performance and has bene®cial effects on performance of growing calves compared to untreated MM. # 2001 Published by Elsevier Science B.V. Keywords: Mustard meal; Glucosinolate; Growth; Body composition; Calves
* Corresponding author. Tel.: 91-1437-20143; fax: 91-1437-20163. E-mail address: [email protected] (M.K. Tripathi).
0377-8401/01/$ ± see front matter # 2001 Published by Elsevier Science B.V. PII: S 0 3 7 7 - 8 4 0 1 ( 0 1 ) 0 0 2 8 6 - 3
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1. Introduction Mustard (Brassica juncea) is the second most important group of oil seeds and India is the second largest producer of rapeseed mustard in the world (Kiresur, 1999), producing 20% of the world's rapeseed mustard. Mustard group oil seed crops have the advantage of being more draught tolerant and disease resistant than other oil seed crops and, with the recent increase in renovated dryland agriculture in India, the production of mustard has been increased substantially. Mustard meal (MM) is the by-product that remains following oil extraction from mustard seeds. On a dry matter (DM) basis (kg 1), MM has 459 g crude protein (CP), 212 g neutral detergent ®ber (NDF) and 128 g acid detergent ®ber (ADF: Newkirk et al., 1997). In spite of its high CP content and well balanced amino acid composition, the use of MM in animal feeding is limited because the meal contains glucosinolate (GLS), compounds that reduce nutritive value and makes MM unpalatable as well as goitrogenic. The GLS degradation products thiocyanate, isothiocyanate and nitriles suppress thyroid uptake of iodine and result in low levels of the thyroid hormones tri-iodothyronine and thyroxin (Barrett et al., 1997). The inclusion of GLS containing meal at higher levels induces metabolic disorders, such as liver and thyroid hypertrophy (Papas et al., 1979; Bell, 1984). These problems are associated with GLS degradation into toxic compounds either by myrosinase inherently present in the cellular compartment of feed or the enzyme present in bacterial micro¯ora (Nugon-Baudon et al., 1990). Most varieties of mustard grown in India contain 12±90 mg g 1 GLS meal (Chauhan et al., 1999). In order to remove or detoxify GLS and improve the nutritional value of rapeseed and mustard meals, acid treatment in combination with heat has been shown to destroy nearly 95% of intact GLS in MM (Tripathi and Agrawal, 1998). Mitigating the deleterious effects of GLS, allow MM to be used in livestock feeding. Our objective was to study the effect of replacing the soybean meal (SBM) with acid treated MM (AMM) and untreated MM (UMM) as protein sources on DM intake, nutrient utilisation, blood biochemical constituents, growth performance, and body composition of calves. 2. Materials and methods 2.1. Acid treatment of mustard meal Untreated MM contained 898.0 g DM, 282.8 g CP, 107.5 g kg 1 ether extract (EE) and 46.2 mg GLS g 1 oil extracted meal. The UMM was treated (Tripathi and Agrawal, 1998) with 16 ml HCl per kg, which raised its moisture content to 40%, after which it was allowed to stand for 72 h at room temperature followed by heating in a forced air oven at 1808C for 2 h. 2.2. Animals and diets Twenty four male crossbred (Jersey Sahiwal) growing calves of 240 15:4 days of age and 87:3 2:52 kg BW were dewormed with `Albendazole'(at 10 mg kg 1 BW) and
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divided into three equal groups based on comparable age and BW. The diet consisted of oat hay (OH) offered ad libitum and a CFM as a supplement to provide adequate protein and other nutrients (ICAR, 1998). The CFM was formulated using SBM or untreated MM or acid treated MM. Untreated MM and acid treated MM completely replaced SBM on a protein basis. Calves were housed individually in stalls and OH and CFM were offered separately. The calves were permitted 10% OH refusals. The daily allowance of CFM was calculated based on previous 14 days OH intake, rate of live weight gain and BW. 2.2.1. Growth trial The growth trial lasted for 16 weeks in a randomised design during which feed intake was recorded daily. Samples of OH and CFM offered were collected once weekly for estimation of DM, and 3 or 4 weeks samples were pooled for chemical analysis. The BW of the calves were recorded for three consecutive days at 14 days intervals before offering feed and water, and mean weights were used to determine growth. 2.2.2. Metabolism trial A metabolism trial was conducted after 40 days of experimental feeding using six calves in each treatment. The metabolism trial lasted for 7 days during which daily intake of OH and CFM, and output of faeces were recorded. Samples of feed offered, feed refusals, and faeces voided were collected every morning. The faeces excreted were collected using total collection bags. Two percent by weight of total faeces daily voided by individual animals was used for DM determination. DM in OH, CFM and faeces were determined by drying at 708C to a constant weight. The dried samples of each day for 7 days collection were pooled, ground through a 1 mm screen and preserved for chemical analysis. Samples of faeces (1/1000) from individual animals were collected every morning for 7 days in a 500 ml Kjeldahl ¯ask containing 25 ml concentrated sulphuric acid for N determination. 2.3. Blood sampling and analysis Blood samples were collected once at the end of the feeding period, through jugular puncture into heparinised tubes for haematological studies and into serum tubes for biochemical assays. Blood glucose, serum total proteins, and albumin were assayed by commercial kits (Span Diagnostics Ltd., New Delhi). Globulin was determined by subtracting albumin from total protein. The haematological measurements (haemoglobin, packed cell volume (PCV), erythrocyte sedimentation rate (ESR), white blood cell (WBC) and red blood cell count (RBC)) were completed by the procedures of Walmsley and White (1994). 2.4. Body composition At the end of the growth trial, animals were weighed for three consecutive days and the average weight was used to determine body composition. Body composition was determined using the antipyrine dilution technique (Brodie et al., 1949) and modi®ed by
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Wellington et al. (1956). Total body water was calculated (Soberman, 1950) from antipyrine level in plasma. The procedures of Bensadoum et al. (1963) for empty BW and Reid et al. (1963) for body fat and body protein estimation were used. The body mineral content was determined by subtracting the water and OM content of the body. 2.5. Chemical analysis Samples of OH, CFM and faeces were analysed for CP, EE and ash as per the procedures, numbered 988.05, 920.05 and 968.08, respectively, of AOAC (1995). The neutral detergent ®bre (NDF) was determined by the procedure of Van Soest et al. (1991) without sodium sulphite or a-amylase, whereas acid detergent ®bre (ADF) and acid detergent lignin (ADL) were determined according to the method described by Goering and Van Soest (1970). Metabolisable energy (ME) was estimated according to Kirchgessner (1995). Total GLS content in UMM, AMM and CFMs was determined using the thymol method (Tholen et al., 1989). Calcium was estimated by the titrimetric method, number 6.011 of AOAC (1965), and phosphorus by a colorimetric method (Donald et al., 1956). 2.6. Statistical analysis Data were analysed using analysis of variance technique (Snedecor and Cochran, 1968). Growth data were analysed by least square analysis (Harvey, 1975), with the model Yij m Ti eij where m is the general mean, Ti the effect of ith treatment (i 1, 3), eij is random error. 3. Results 3.1. Glucosinolate content of feeds The GLS content of UMM was 46.2 mg g 1 meal, which was reduced to 4.5 mg g 1 in AMM (Table 1). The GLS was 2.6 and 26.4 mg g 1 DM in CFM containing AMM and UMM, respectively. 3.2. Growth trial The BW gain was 56.6, 57.4 and 30.6 kg, respectively, in calves fed the diet with SBM, AMM and UMM and these differences were signi®cant (P < 0:05). Total BW gain and ADG was lower (P < 0:01) 26.0 kg and 220 g, respectively, in calves fed the diet with UMM compared with SBM or AMM calves. However, ADG, feed intake and FCR did not differ between SBM and AMM fed calves (Table 2). Feed intake was higher (P < 0:05) in SBM and AMM fed calves compared to UMM calves whereas FCR was lower in UMM fed calves. Calves fed the diet with UMM consumed 5.4±6.3 kg more (P < 0:01) feed for each kg gain to those fed the SBM or AMM diet, respectively.
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Table 1 Ingredient and chemical composition of compound feed mixtures (CFMs) of three treatments and hay used for experimental feeding Protein source in compound feed mixture Soybean meal
Oat hay
Mustard meal Acid treated
Untreated
1
Ingredient composition (g kg ) Soybean meal Acid treated mustard meal Untreated mustard meal Deoiled rice bran Wheat grain Salt Mineral mixturea
265 ± ± 350 355 10 20
± 495 ± 240 235 10 20
± ± 495 240 235 10 20
Chemical composition (g kg 1 of DM) Organic matter Crude protein Ether extract Neutral detergent fiber Acid detergent fiber Acid detergent lignin Calcium Phosphorus ME (MJ kg 1 DM)b Glucosinolate (mg g 1 DM)
860.5 216.3 15.2 509.0 167.0 35.5 17.0 12.4 11.06 NDc
870.3 207.5 16.2 499.0 166.0 47.0 20.0 11.6 11.48 2.6
871.8 216.0 16.9 481.0 168.0 50.2 22.0 11.5 11.78 26.4
883.5 147.5 20.0 609.5 388.5 57.0 16.5 1.6 9.02 NDc
a Composition: calcium 320 g kg 1, phosphorus 62 g kg 1, manganese 2.7 g kg 1, zinc 2.6 g kg 1, iron 1000 ppm, fluorine 900 ppm, iodine 100 ppm, copper 100 ppm. b Estimated according to Kirchgessner (1995). c Not detected.
Table 2 Growth performance of calves on the three treatmentsa Protein source in compound feed mixture Soybean meal
Initial body weight (kg) Final body weight (kg) Average daily gain (g per day) Total feed intake (kg) Feed conversion ratio (g/g) a
88.6 145.2 a 472 a 575 a 10.15 a
S.E.M.
Mustard meal Acid treated
Untreated
87.2 148.6 a 495 a 549 a 9.24 a
87.9 118.5 b 255 b 476 b 15.56 b
Values having different letters in rows differ significantly (P < 0:05).
5.3 6.8 57 21 1.04
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Table 3 Nutrient intake, digestibility and N utilisation of three diets used in metabolic triala Protein source in compound feed mixture Soybean meal
S.E.M.
Mustard meal Acid treated
Untreated
3.60 a 1.19 4.79 4.06 a 133.6 a
3.19 b 1.18 4.37 3.93 b 127.7 b
2.96 b 1.17 4.13 3.88 b 122.4 b
0.13 0.04 0.41 0.05 1.4
Metabolisable energy intake MJ per day
45.63
42.37
40.48
3.43
Digestibility (%) Dry matter Organic matter Crude protein Neutral detergent fiber Acid detergent fiber
63.1 65.1 65.3 61.3 50.7
61.8 63.7 66.9 58.6 45.6
62.5 64.4 67.6 57.8 49.8
1.6 1.2 1.3 1.4 2.0
Nitrogen utilisation Intake (g per day) Faecal (g per day) Absorbed (g per day) Absorbed (% intake)
128.5 a 45.3 83.2 a 64.7
121.9 b 40.6 73.9 b 64.5
108.5 b 34.1 74.4 b 68.6
2.0 4.4 2.2 1.9
Dry matter intake (kg per day) Hay (kg per day) Concentrateb (kg per day) Total (kg per day) (% BW) (g kg 1 W0.75)
a b
Values having different letters in rows differ significantly (P < 0:05). The relevant concentrate for the treatment (Table 1).
3.3. Metabolism trial Intake of DM in CFM, and in total, and ME intake were similar among the three groups (Table 3). The DMI of OH was higher (P < 0:01) in SBM fed calves compared to those fed AMM or UMM diets. 3.3.1. Nutrient digestibility and nitrogen utilisation Digestibility of DM constituents and ®ber fractions did not differ among diets. The N intake was lowest (P < 0:05) in calves fed the UMM diet compared to those fed SBM or AMM. The N voided in faeces showed the same trends as N intake. Absorption of N (g per day) was highest (P < 0:05) in calves fed SBM diet. 3.4. Blood biochemical constituents and body composition The albumin concentration was lowest in calves fed the UMM diet. Whereas, serum protein and blood glucose did not differ among diets. The ESR was highest in UMM, intermediate in AMM and lowest in SBM fed calves, Other haematological and body (i.e. water, protein, fat and mineral) constituents did not differ among treatment (Table 4).
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Table 4 Blood biochemical and haematological constituents, and body composition of calvesa Protein source in compound feed mixture Soybean meal
S.E.M.
Mustard meal Acid treated
Untreated
1
Blood constituents (mg dl ) Total serum protein Albumin Globulin Blood glucose
7.79 4.54 a 3.25 70
8.56 4.93 a 3.63 73
6.73 2.79 b 3.94 71
13.84 0.68 0.28 <1
Haematological constituents Haemoglobin (g%) Packed cell volume (%) Erythrocyte sedimentation rate (%) Red blood cell counts (106 mm 3) White blood cell counts (103 mm 3)
7.7 23.8 3.5 c 3.8 11.0
7.2 22.3 5.8 b 3.8 11.0
7.4 26.7 7.8 a 4.1 10.0
0.3 1.1 <0.1 0.2 0.5
Body composition (g kg Water Protein Fat Mineral matter a
1
empty body weight)
554 157 226 61
456 138 348 58
596 168 186 50
34 10 42 5
Values having different letters in rows differ significantly (P < 0:05).
4. Discussion Hydrolysis of GLS is caused by myrosinase enzyme, which is present in MM. Myrosinase expresses maximum activity at 15±40% moisture level (Mukherjee et al., 1976) and 3.5±8.0 pH (Gil and Macleod, 1980). In the present experiment, acid treatment of MM reduced GLS content sharply (90.3% GLS hydrolysed). The HCl treatment applied to MM increased the moisture level to 40% and changed the pH to favour catalytic activity of the myrosinase enzyme required for GLS degradation. Added HCl reduced the pH of the meal from 5.35 to 2.79 (Tripathi and Agrawal, 1998), which altered the nature of GLS degradation products. The heat applied during acid treatment (1808C for 2 h) also destroyed GLS and/or some of the GLS degradation products evaporated. Digestive processes in the rumen are the result of microbial and fungal activities, which are reduced due to the antifungal and antimicrobial activities of GLS and/or GLS degradation products (Manici et al., 1997). In contrast to this study, lower digestibility of DM constituents and ®ber fractions were reported in RSM (Sharma et al., 1980; Lardy, 1993) and MM (Tripathi et al., 2001a) diets. The metabolic trial in this study was conducted after only 40 days of experimental feeding. Possibly due to this short adaptation period, the known deleterious effects of GLS were less pronounced on the digestive ability of the calves, and so treatment differences failed to reach to statistical signi®cance. Tripathi et al. (2001b) reported that dietary glucosinolates exhibit deleterious effects on the digestive ability of calves, whereas glucosinolates require some time to manifest its
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deleterious effect on digestibility and growth. Lower DMI of calves fed untreated MM, compared to SBM or acid treated MM, was probably due to depressed appetite associated with the GLS or its degradation products (Pusztai, 1989; Duncan and Milne, 1990). Lower feed intake was also reported by Olsson (1978) and Tripathi et al. (1996, 1997) on RSM diets compared to SBM diets. Absorption of N containing compounds on acid treated MM diets might be higher in the small intestine because the heat applied during acid treatment of MM reduces degradability of protein in the rumen (érskov, 1981) thereby improving ef®ciency of amino acid absorption in the small intestine (Mustafa et al., 1999). The higher growth rate, better FCR and higher albumin concentration in calves fed the acid treated MM diet are probably the combined effects of higher N utilisation and negligible amounts of GLS. Lower albumin concentrations in untreated MM fed calves was probably the effect of reduced liver synthesis (Eggum, 1989; Walmsley and White, 1994; Rosenthal, 1997) due to the deleterious effects of GLS on liver function causing reduced N utilisation and poor growth. 5. Conclusion Mustard (B. juncea) meal, incorporated in diets of crossbred growing calves as a replacement for SBM negatively affected DM intake, growth rate, feed conversion ratio, N utilisation and serum albumin concentrations, presumably due to high the GLS content. However, when MM was treated with HCl, it could replace SBM without negative effects on N utilisation and growth performance. Acknowledgements The author wish to thank Director, Central Sheep and Wool Research Institute, Avikanagar for study leave and Dr. Mahender S. Rahal, Professor (Animal Nutrition), Department of Animal Science, Govind Ballabh Pant University of Agriculture and Technology, Pantnagar, for critical suggestions and cooperation during the experiment. References AOAC, 1965. Official Methods of Analysis, 10th Edition. Association of Official Analytical Chemists, Washington, DC. AOAC, 1995. Official Methods of Analysis, 16th Edition. Association of Official Analytical Chemists, Washington, DC. Barrett, J.E., Klopfenstein, C.F., Leipold, H.W., 1997. Detoxification of rapeseed meal by extrusion with an added basic salt. Cereal Chem. 74, 168±170. Bell, J.M., 1984. Nutrients and toxicants in rapeseed meal. A review. J. Anim. Sci. 4, 996±1010. Bensadoum, A., Van Niekerk, B.D.H., Paladines, D.L., Reid, J.T., 1963. Evaluation of antipyrine, N-acetyl-4aminoantipyrine and shrunk body weight in predicting the chemical composition of sheep body. J. Anim. Sci. 22, 604±612. Brodie, B.B., Axelrod, J., Soberman, R., Levy, B.B., 1949. The estimation of antipyrine in biological materials. J. Biol. Chem. 179, 25±29.
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