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Effect of substitution of soybean meal by detoxified karanja cake on diet digestibility, growth, carcass and meat traits of sheep
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Effect of substitution of soybean meal by detoxified karanja cake on diet digestibility, growth, carcass and meat traits of sheep S.B.N. Rao a,∗ , D. Dinesh Kumar b,1 a
Animal Nutrition Division, National Institute of Animal Nutrition and Physiology, Adugodi, Bangalore 560 030, India CNPq-TWAS Research Scholar, Laboratory of Animal Nutrition, Center for nuclear energy in Agriculture (CENA), University of Sao Paulo, Brazil b
a r t i c l e
i n f o
Article history: Received 17 July 2013 Received in revised form 12 March 2015 Accepted 12 March 2015 Available online xxx Keywords: Biochemical metabolites Carcass traits Digestibility Karanja cake Sheep
a b s t r a c t The non-conventional karanja cake is rich in protein (around 30% CP) and can be used in livestock feed as a protein source instead of conventional protein supplement cake like soybean meal (SBM), groundnut cake, etc. The present study was carried out to research the effect of partially substituted soybean meal with detoxified karanja cake (dKC) on performance of ram lambs. Twenty-four ram lambs were randomly divided into four groups (n = 6) and fed different levels (%) of detoxified karanja cake (0% replacement, control; 25% replacement, dKC-25; 50% replacement, dKC-50 and 75% replacement, dKC-75) in concentrate mixtures for 140 days. dKC was incorporated in the concentrate mixtures at the expense of soybean meal, maize grain and wheat bran at 9, 18 and 29% in dKC-25, dKC-50 and dKC-75, respectively on fresh basis. As the level of karanja in the diet increased, DMI was found to be decreasing significantly. Similar to these effects, N-retention was reduced leading to significant reduction of body weight in high karanja cake replaced groups. Similar trend was observed in OM, CP, and ADF digestilities and reason attributed to increased dietary level of karanja cake. However, detrimental effects were not observed on the levels of total protein, albumin, globulin, serum urea, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) indicating residual ANF present in dKC did not exert any adverse effects. The effects on hot carcass weight, weights of liver and testes are following a decreasing trend while that weight of kidney is increasing with level of karanja in the diet. Our findings highlights that the detoxified karanja cake can be added as replacement of soybean meal (SBM) at low levels. However, higher levels of replacement (above 9 per cent of concentrate mixture) warrant caution due to its adverse effect on studied parameters. © 2015 Elsevier B.V. All rights reserved.
1. Introduction Karanja (Pongamia spp.) is grown in parts of humid tropical regions of Asia and Oceania. The seed contains 20–30%
∗ Corresponding author. Tel.: +91 8025711304; fax: +91 8025711420. E-mail addresses: [email protected], [email protected] (S.B.N. Rao). 1 The second author is at present pursuing his Ph.D. in Brazil. The reported work was conducted in Bangalore. He worked as Research Fellow in the project.
protein, 27–39% oil and a group of furano-flavonoids that constitutes 5–6% by weight of the oil (Bringi and Mukerjee, 1987). In a recent study, large variations were observed in seed size, composition in terms of protein and fat, incriminating factors like karanjin, pongamol and trypsin inhibitor activity (TIA) collected from different regions of Karnataka (Dineshkumar et al., 2011). The seed kernel after extraction of oil is rich in protein (around 30% CP) which can be used in animal and poultry feed as a protein source (Konwar et al., 1987; Vinay and Kanya, 2008). However, raw expeller karanja cake (rKC) is not commonly used as
http://dx.doi.org/10.1016/j.smallrumres.2015.03.010 0921-4488/© 2015 Elsevier B.V. All rights reserved.
Please cite this article in press as: Rao, S.B.N., Kumar, D.D., Effect of substitution of soybean meal by detoxified karanja cake on diet digestibility, growth, carcass and meat traits of sheep. Small Ruminant Res. (2015), http://dx.doi.org/10.1016/j.smallrumres.2015.03.010
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a feed for livestock due to its poor palatability and presence of antinutritional factors (ANFs), i.e. furanoflavones like karanjin, pongamol and certain other polyphenolic compounds (Bringi and Mukerjee, 1987; Prabhu et al., 2002) leading to depression in performance of livestock (Srivastava et al., 1990; Nagalakshmi et al., 2011; Panda et al., 2004). Various methods, namely refluxing with 2% HCl (Mandal, 1985) water leaching, solvent extraction, acid and alkali treatment, autoclaving were adopted to improve palatability and nutritive value of detoxify karanja cake. Among all the methods, solvent extraction was found to be more efficient to remove karanjin (Prabhu et al., 2002) and is being commonly used as a method to detoxify karanja cake. Singh et al. (2006) reported that long term feeding of expeller karanja cake or solvent extracted karanja cake had deleterious effects on the nutrient utilization, blood biochemical profile, rumen fermentation pattern, carcass characteristics and manifested clinico-pathological changes in bone and tissues of vital organs. Further studies indicated that inclusion of expeller karanja cake at 12% in complete diets constituting 390 mg karanjin per kg depressed the performance in lambs (Nagalakshmi et al., 2011). Keeping in view the adverse effects exerted by feeding of karanja cake, present study aimed to fill the gaps in scale up of detoxification methodology of karanja cake at industrial scale and its possible utilization in livestock rations replacing soybean cake. The effects on physiological responses in terms of gene expression of LH receptor and IGF-I genes related to testicular function, testicular architecture and hormones was already reported (Dineshkumar et al., 2013). The objective of the particular experiment is to test different levels of detoxified karanja cake in rations of growing lambs based on its effects on intake, digestibility, growth, carcass and meat traits. 2. Materials and methods 2.1. Source of detoxified karanja seed cake (dKC) Bulk production of detoxified meal required for feeding trial was collected from industrial partners (M/s Ayurvet Pvt. Limited, Baddi, Himachal Pradesh, India and M/s Ganesh Scientific Research Foundation, New Delhi,
India). A process was developed to detoxify karanja seeds initially by crushing and removing the oil using hexane. Then the meal was further detoxified by soaking the seeds in aqueous methanol mixture and drying. 2.2. Feed preparation The requirements of crude protein (CP) and total digestible nutrients (TDN) suggested by Indian Council of Agricultural Research (ICAR, 1998) formed the guidelines for feeding lambs. The required quantities of feed ingredients like finger millet (Eleusine corocana) straw, maize (Zea mays) grain, soybean (Glycine max) meal, wheat (Triticum aestivum) bran, mineral mixture and salt were procured from commercial source for the entire duration of the experiment Maize grain and soybean meal were ground in hammer mill and mixed with wheat bran, mineral-vitamin premix and salt in horizontal mixer. Four iso-nitrogenous and iso-caloric concentrate mixtures were prepared by making minor adjustments in the proportions of wheat bran and maize grain were made in the formulation (Table 1). 2.3. Experimental animals and housing 24 ram lambs were used in this experiment (body weight: 13.7 ± 0.5 kg; aged around 6 months) for a period of 140 days in a permanent open type shed having provision for tying the animals in separate enclosures and also feeding trough. Ram lambs were fed individually and reared under hygienic and uniform managerial conditions throughout the experiment. Ram lambs were offered clean water 2–3 times daily for the entire duration. They were dewormed using broad spectrum anthelmentic (albendazole®) at the rate of 10 mg per kg body weight twice at 21 days interval at the beginning of the trial and all the animals were confirmed to be parasite free by faecal examination. Ram lambs were randomly divided into four groups (n = 6) and were offered concentrate mixture incorporating dKC as a replacement of SBM. Different levels (%) of karanja cake were fed (0% replacement, control; 25% replacement—9% dKC in concentrate mixture on fresh basis, dKC-25; 50% replacement—18% dKC in concentrate mixture on fresh basis, dKC-50 and 75% replacement—29% dKC in concentrate mixture on fresh basis, dKC-75). dKC was incorporated in concentrate mixtures (on fresh basis) at 9, 18 and 29% in dKC-25, dKC-50 and dKC-75, respectively. The lambs were offered the respective concentrate mixtures daily between 9:00 and 9:30 h, to meet protein requirements (ICAR, 1998) for maintenance and expected daily gains of 75 g. The lambs were fed concentrate mixture around 1.6–1.7 percent of bodyweight. The remaining requirements were met through feeding of chaffed finger millet (Eleusince corocana) straw (∼1.6% of body weight), which was offered after 2:00 h of feeding concentrate mixture fed to ensure its maximum consumption. On an average lambs consumed around 250–350 g each of concentrate and ragi straw across different treatment groups. All lambs were weighed individually at weekly intervals to assess body weight changes and growth
Table 1 Ingredients and chemical composition of feeds used in the experiments. Item
dKC 0
dKC 25
dKC 50
dKC 75
Ragistraw
Ingredients (%) Maize grain Soybean meal Karanja detoxified cake Wheat bran Mineral mixtureA Salt Dry matter (%) Organic matter (% DM) Ether extract (% DM) Crude protein (% DM) Acid detergent fibre (% DM) Karanjin (%)a Trypsin inhibitor activity (g/g)a
31 30 0 36 2 1 96.84 91.70 2.61 22.75 8.89 – –
31 22.5 9 34.5 2 1 97.26 92.13 3.00 22.57 8.63 0.0027 35.1
33 15 18 31 2 1 97.51 93.28 3.73 22.30 9.05 0.0054 70.2
34 7.5 29 26.5 2 1 97.01 93.69 3.35 21.72 8.57 0.0087 113.1
93.53 91.58 1.18 4.05 43.35 – –
dKC-0, detoxified karanja cake 0% replacement with Soybean cake; dKC-50, detoxified karanja cake 50% replacement with Soybean cake and dKC-75, detoxified karanja cake 75% replacement with Soybean cake. A Commercial mineral mixture with the composition per kg: Cobalt-150 mg, copper-1200 mg, iodine-325 mg, iron-5000 mg, magnesium-6000 mg, potassium-1500 mg, selenium-100 mg, sodium-10 mg, sulphur-5.9 mg, zinc-0.942%, DL Methinonine-9600 mg, L-lysine, mono hydrochloride-4400 mg, calcium-24%, phosphorous-12% was used. a Calculated values on the basis of concentrations of detoxified karnaja cake.
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2.4. Metabolism trial After 90 days of the experimental feeding, a 6-days metabolism trial was carried out on all lambs (n = 24) involving total faecal and urine collection. This collection is preceded by 2 days adaptation period for lambs’ acclimatization to faecal and urine bags. During the trial period, all the lambs were tied with cotton ropes. The faeces collection was facilitated by tying tarpaulin faecal bags and urine was collected in polythene bags. The contents of faeces and urine from bags were cleared at regular intervals manually in the airtight plastic containers contained known quantity of H2 SO4 . Daily feed offered, residues left and faeces and urine excreted was recorded on 24-h basis. Feeds, refusals and an aliquot of faeces were dried at 100 ± 5 ◦ C for 24 h for the determination of DM. The dry samples were ground to pass through a 1-mm sieve and pooled for proximate analysis. Day to day fresh samples of faeces (1/10) was preserved with 1:4 sulphuric acid for N estimation. 2.5. Collection and analysis of rumen liquor and blood samples After the metabolism trial, rumen liquor was collected from all the sheep with the help of stomach tube. Little suction was applied to collect the rumen liquor from the sheep (Rao et al., 2012). The pH of rumen liquor was recorded immediately after collection using a pre-calibrated digital pH Meter. The strained rumen liquor was analyzed for ammoniaN (Conway, 1957), total-N and trichloroacetic acid precipitable-N (TCA ppt.-N) (AOAC, 2000). Blood was collected from jugular vein. Serum was separated and stored at −80◦ C till analyzed. Biochemical profiles such serum protein (g/dl), albumin (g/dl), glucose (g/dl), serum urea (mg/dl) and enzymes such as alanine amino tranferease (ALT) and aspartate aminotransferase (AST) (IU/l) were estimated using biochemical kits purchased from Span Diagnostics Limited, Surat, Gujarat, India. The globulin concentration was measured as difference between total protein and albumin. 2.6. Laboratory analysis Proximate analysis of the concentrate mixtures, finger millet straw and faeces viz. CP, EE and ash were determined by the methods of AOAC (2000). Acid detergent fibre (ADF) was estimated using the procedure of Vansoest et al. (1991). The karanjin was estimated by separation of components by TLC on pre-coated silica gel G 60 F254 plates developed on toluene: ethyl acetate (7:3 v/v) and detection at 260 nm in absorbance mode (Ravikanth et al., 2009) and Trypsin inhibitory activity (TIU/g) indirectly by inhibiting the activity of trypsin. A synthetic substrate Nu-benzoyl OL-arginine p-nitroanilide (BAPNA) is subjected to hydrolysis by trypsin to produce yellow coloured p-nitroanilide. The degree of inhibition by the extract of the yellow colour production is measured at 410 nm (Chitra and Sadasivam, 1986). 2.7. Slaughter and carcass evaluation Feed was withheld overnight with free access to water and animals were slaughtered in a local abattoir by Halal method (Gracey, 1981). After slaughter, the head and feet were removed and the animals were partially skinned lying on their back on the floor. Thereafter, the animals were suspended by the hind legs for further skinning. Carcass and noncarcass components were weighed immediately after slaughter. Lungs, trachea and heart were weighed as one piece and designated as pluck. Non-carcass components included head, skin, feet, digestive tract, liver, spleen, pancreas and pluck. Weight of digestive contents was computed as the difference between full and empty digestive tract. The empty live weight was computed as the difference between slaughter weight and weight of digesta content. 2.8. Meat quality evaluation The longissimus dorsi muscle was collected, within 20 min of slaughter, trimmed of fat and frozen at −80 ◦ C till analyzed for meat quality traits. The pH of minced L. dorsi muscle was recorded by using a digital pH meter. Proximate composition of longissimus dorsi was determined on muscle
800.00 Dry maer intake (g/day)
rate. The individual intake of nutrients such as DM, CP, DCP and TDN per unit gain was calculated by dividing intake of nutrients obtained from individual animals during sub trial for arriving values of nutrient conversion efficiency.
3
700.00 600.00 500.00 400.00 300.00 200.00 y = -12.266x + 688.62 R² = 0.985
100.00 0.00 0
2
4
6
8
10
12
14
16
Level (%) of Karanja cake in the diet DMI (g/day)
Linear (DMI (g/day))
Fig. 1. Regression analysis of average dry matter intake (Y-axis) and percent of Karanja cake in the diets.
samples according to AOAC (2000). The sensory evaluation of pressurecooked meat samples for colour, odour, juiciness, tenderness and overall palatability was performed by a taste panel consisting of 10 panelists using 9 point scale (9 = like extremely, 1 = dislike extremely).
2.9. Statistical analysis Individual sheep data was considered as a replicate and a one-way analysis of variance (ANOVA) was done using completely randomized design (Snedecor and Cochran, 1994) by a mathematical model of Harvey (1975). SPSS is a software package used for statistical analysis (SPSS Inc., Chicago, IL, USA) for analysis of variance assuming for independent constant variance structure with post-hoc tukey to find the pair wise significance between treatments. A P-value of less than or equal to 0.05 was accepted to indicate statistical significance. Repeated measure analysis was performed on for body weights recorded at different periods (weeks) using JMP® 11.2.0 software by fitting a model with a role variable as body weights, constructing model effects (treatment and period) and interactions (Treatment × Period) and generated least square means were subjected for significance using tukey post-hoc. Linear regression analysis was performed on different parameters such as dry matter intake, N excretion (faeces) and urine, N retained, hot carcass weight, organ weighs (y variable) with % of Karanja in the diet to construct a linear bivariate fit.
3. Results 3.1. Chemical composition of concentrate mixtures CP content (%) of concentrate mixtures was found to be 21.7–22.8 in all groups (Table 1). EE% was found to be ranging from 2.6 in control to 3.7 in dKC-50. Increased EE in case of dKC groups could be due to more EE content of dKC used in the experiment (3.85%). dKC had shown residual concentration of Karanjin, 0.03% W/W and TIA, 390 g/g. 3.2. Intake, digestibility and balance of nitrogen The daily dry matter intake of lambs was regressed against per cent of incorporation of Karanja in the diet indicated significantly negative correlation (Fig. 1) indicating as the level of karanja increased in the diet the DMI is following a decreased trend. This was also reflected on reduced DCP intake (g/d) in dKC-50 and dKC-75 groups compared to control and dKC-25. Compared to control, dKC-50 and dKC-75 groups had low digestibilities of DM, OM, ADF and T-CHO. CP digestibility was lowest in dKC-75 (Table 2).
Please cite this article in press as: Rao, S.B.N., Kumar, D.D., Effect of substitution of soybean meal by detoxified karanja cake on diet digestibility, growth, carcass and meat traits of sheep. Small Ruminant Res. (2015), http://dx.doi.org/10.1016/j.smallrumres.2015.03.010
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4 Table 2 Intake, digestibility in different groups. Attributes
Diet
SEM
dKC 0
dKC 25
dKC 50
dKC 75
Dry matter intake (g/d) DOM intake (g/d) DCP intake (g/d) TDN intake (g/d)
684.81 437.04 73.02b 436.50a
643.92 407.84 69.36b 411.49a,b
565.16 346.63 57.28a,b 350.71a,b
510.75 317.13 46.17a 315.92b
Nutrient digestibility (%) Organic matter Crude protein ADF
69.71b 79.29b 47.66b
68.97a,b 79.79b 44.62b
66.25a 75.68a,b 37.57a
26.09 17.49 3.25 17.31
66.68a,b 68.97a 37.61a
0.49 1.79 1.31
dKC-0, detoxified karanja cake 0% replacement with Soybean cake; dKC-50, detoxified karanja cake 50% replacement with Soybean cake and dKC-75, detoxified karanja cake 75% replacement with Soybean cake. Means bearing different superscripts (a and b) in a row differ significantly (P < 0.05).
groups (Table 3). Differences in total protein, albumin, aspartate aminotransferase (AST), alanine aminotransferase (ALT) (IU/l) and serum urea (mg/dl) were observed to be same in all groups indicating the residual ANF’s (Karanjin, pongamol, TIA and others) have not exerted any systemic effects in elevating or affecting any of the profiles studied. 3.5. Carcass characteristics, composition of Longissmus dorsi and organo-leptic evaluation of meat Fig. 2. Bodyweight least square means (kg) across different treatments (1, dKC-0, detoxified karanja cake 0% replacement with Soybean cake 19.34; 2, dKC-25, detoxified karanja cake, 18.64; 3, dKC, 50, 50% replacement with Soybean cake, 17.02; and 4, dKC-75, detoxified karanja cake 75% replacement with Soybean cake 15.52 kg).
3.3. Body weights during experimental period
Hotcarcass weight was following declining trend as level of karanja cake increased in diet (Fig. 3). Similar to hot carcass weight, liver, heart and testes weights following declining trend. However, weight of kidney was following a increasing trend. Analyzed parameters like dry matter, fat, protein and ash were found to be similar in all treatment groups indicating no adverse effect of feeding dKC on chemical composition of meat. Organoleptic evaluation in terms of like appearance, colour, texture and consistency, aroma and flavor and overall acceptability were found to be same in all tested groups (Table 4).
The least square means of body weights across different treatments presented (Fig. 2). The average body weights recorded across different treatments were similar only up to 25% of replacement. At higher levels of replacements (50 and 75%), body weights were significantly (P < 0.05) reduced.
4. Discussion
3.4. Rumen metabolites and serum biochemical profiles
4.1. Chemical composition of concentrate mixtures
Rumen fermentation profiles such as pH, ammonia-N, TCA-ppt N, total-N were similar (P > 0.05) in all treatment
Karanjin contents of expeller/solvent extracted Karanj seed cake reported by various workers 0.32% by
Table 3 Rumen metabolites, serum biochemical profiles of lambs. Attributes
pH Ammonia nitrogen (mg/100 ml SRL) Total nitrogen (mg/100 ml SRL) TCA ppt nitrogen (mg/100 ml SRL) Serum protein (g/100 ml) Serum albumin (g/100 ml) Urea (mg/dl) Aspartate aminotransferase (IU/l) Alanine aminotranferease (IU/l)
Diet
SEM
dKC 0
dKC 25
dKC 50
dKC 75
6.46 40.61 101.62 47.37 7.00 4.83 60.10 143.6 38.61
6.44 45.29 100.28 50.96 6.93 4.91 58.70 140.6 34.10
6.42 35.93 99.17 56.66 6.97 4.89 50.45 147.8 36.86
6.40 33.88 99.54 60.48 6.40 4.48 56.24 145.8 30.97
13.27 2.29 2.67 0.30 0.57 0.11 1.95 9.51 3.16
dKC-0, detoxified karanja cake 0% replacement with Soybean cake; dKC-50, detoxified karanja cake 50% replacement with Soybean cake and dKC-75, detoxified karanja cake 75% replacement with Soybean cake. Means bearing different superscripts (a and b) in a row differ significantly (P < 0.05).
Please cite this article in press as: Rao, S.B.N., Kumar, D.D., Effect of substitution of soybean meal by detoxified karanja cake on diet digestibility, growth, carcass and meat traits of sheep. Small Ruminant Res. (2015), http://dx.doi.org/10.1016/j.smallrumres.2015.03.010
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Table 4 Caracass characteristics, composition of Longissmus dorsi and organo-leptic evaluation of meat by semi-trained panelists. Attributes
Diet
SEM
dKC-0
dKC-25
dKC-50
dKC-75
Live body weight (kg) after 12 h of fasting Hot carcass weight (kg)
24.08b 11.04b
24.75b 11.10b
22.33a,b 9.84a,b
17.75a 8.05a
0.93 0.43
Weight of vital organs (g) Liver Kidneys Heart Testes Spleen
391b 57 122 172 51
384b 60 144 250 57
325a,b 61 140 202 59
271a 68 107 164 48
16 3 6 33 3
Chemical composition (% on fresh basis) Dry matter (%) Total fat (%) Ash (%) Protein (%) pH
27.30 5.22 1.01 19.82 5.65a
28.32 5.28 1.06 21.17 5.73b
25.91 3.61 1.10 20.47 5.66a
27.63 4.16 1.15 21.93 6.05c
0.45 0.28 0.02 0.35 0.04
Organoleptic evaluation Appearance Colour Texture and consistency Aroma and flavour Overall acceptability
6.63 6.63 5.50 5.94 6.38
6.13 6.13 6.13 5.88 6.38
5.63 5.63 6.38 5.94 6.50
– – – – –
0.27 0.27 0.26 0.21 0.26
g/day
dKC-0, detoxified karanja cake 0% replacement with Soybean cake; dKC-50, detoxified karanja cake 50% replacement with Soybean cake and dKC-75, detoxified karanja cake 75% replacement with Soybean cake. Means bearing different superscripts (a,b,c) in a row differ significantly (P < 0.05).
4.50 4.00 3.50 3.00 2.50 2.00 1.50 1.00 0.50 0.00
to this, Makkar et al. (1990) have reported lower total tannins and condensed tannins content in karanj cake and protein precipitation was not reported. The calculated concentrations of karanjin and TIA in the present study have increased linearly as level dKC increased in the concentrate mixtures (0.0027–0.0087 for karanjin and 35.1 and 113.1 for TIA).
y = -0.0802x + 3.9291 R² = 0.8527
0
2
4
6
8
10
12
14
16
Level (%) of Karanja cake in the diet
g/day
N- excreon (Urine)
Linear (N- excreon (Urine))
9.00 8.00 7.00 6.00 5.00 4.00 3.00 2.00 1.00 0.00
y = -0.2241x + 8.075 R² = 0.9818 0
2
4
6
8
10
12
14
16
Level (%) of Karanja in the diet N-retenon
Linear (N-retenon )
Fig. 3. Regression analysis of N-excretion (Urine) and N-retained (Y-axis) against percent of Karanja cake in the diets.
Nagalakshmi et al. (2011), 0.32% by Panda et al. (2006), 0.19% by Prabhu et al. (2002) and 0.29% by Soren et al. (2007). Vinay and Kanya (2008) have reported 0.94% of tannins along with other anti-nutritional factors, such as phytate (0.65%) and trypsin inhibitors (39 IU/mg). Contrary
4.2. Intake, digestibility and balance of nitrogen Many studies involved with feeding of either solvent extracted karanja cake or expeller karanja cake were reported (Konwar et al., 1987 in crossbred bull calves; Srivastava et al., 1990 in growing kids; Singh et al., 2006; Ravi et al., 2000; Prabhu, 2002; Soren and Sastry, 2009; Nagalakshmi et al., 2011 in growing lambs). As observed in present study reduction in feed intake was observed in cross-bred bulls fed 26.5% deoiled karanja cake based concentrate mixture (Konwar et al., 1987) and in lambs fed with concentrate mixtures containing either 24% expeller karanja cake or 20% deoiled karanja cake (Singh et al., 2006) or 22.5% processed deoiled karanja cake (Soren and Sastry, 2009) or 12% expeller karanja cake (Nagalakshmi et al., 2011). On the contrary, no effect on DMI has been reported in goat kids receiving deoiled karanja cake at 40% level in concentrate mixture (Srivastava et al., 1990). Other workers (Ravi et al., 2000; Prabhu, 2002) also reported similar observations on dry matter intake in growing lambs receiving de-oiled karanja cake at 20% level. Similar to our results, reduction in nutrient digestibility was observed (Srivastava et al., 1990) in goat kids receiving 12% de-oiled karanja cake and 24% expeller karanja cake in lambs by Ravi et al. (2000). However, no effect on DM, OM digestibilities (Soren
Please cite this article in press as: Rao, S.B.N., Kumar, D.D., Effect of substitution of soybean meal by detoxified karanja cake on diet digestibility, growth, carcass and meat traits of sheep. Small Ruminant Res. (2015), http://dx.doi.org/10.1016/j.smallrumres.2015.03.010
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4.3. Body weights during experimental period The results obtained in the present experiment are consistent with earlier similar experiments involved with feeding karanja cake (Ravi et al., 2000; Soren et al., 2008; Nagalakshmi et al., 2011) in growing lambs fed either expeller or processed karanja cake. 4.4. Rumen metabolites and serum biochemical profiles In line with the current study, Singh et al. (2006) found no significant differences in pH, NH3 -N, total-N and TCA ppt.-N in the rumen liquor of lambs fed expeller pressed (24.0%) and solvent extracted karanj cake (20.0%) for a period of 255 days. Differences in total protein, albumin,
14.00 12.00
kg
10.00 8.00 6.00 4.00
y = -0.2121x + 11.519 R² = 0.8822
2.00 0.00
5
0
10
15
20
Level (%) of Karanja in the diet Hot carcass weight (Kg)
Linear (Hot carcass weight (Kg))
80.00 75.00 y = 0.3696x + 58.414 R² = 0.5624
70.00 Gram
and Sastry, 2009) and nutrient retention (Panda et al., 2004) was observed with incorporation of raw or processed expeller or solvent extracted karanja cake in lambs and broilers, respectively. The negative effects of karanja cake feeding on intake, digestibility and subsequent nutritive value of diet mainly because of residual ANFs like karanjin, trypsin inhibitors, pongamol, pongapin and lanceolatin B present in residual oil portion of karanja cake which might have affected the digestibility of these nutrients because karanjin possesses the insecticidal and antibacterial properties and also phenolic compounds which are well known for depressing the nutrients digestibility (Rangasamy and Seshadri, 1940; Roy et al., 1977). The reduction in protein digestibility observed the present study might be due to residual tryspin inhibitors and tannins present in karanja cake. The adverse effect of trypsin inhibitors is mainly on the pancreas, which responds to the inhibitors by enhanced synthesis and secretion of proteolytic enzymes. The pancreatic enzyme secretion is regulated by a negative feedback mechanism mediated by intestinal trypsin and chymotrypsin, and complex formation of trypsin with the inhibitors leading to a reduction of free trypsin in the small intestine (Alumot and Nistan, 1961). This reduction activates the pancreas-stimulating hormone, cholecystokinin, the release of which from the intestinal mucosa is inhibited by free trypsin (Wilson et al., 1978). Regression analysis of N-retained (g/day) with % dietary incorporation of dKC indicated amount of N-retained is linearly decreased as the level of dKC increased in the diet. This is primarily due to low intake and subsequent urinary excretion of nitrogen (Fig. 4). However, faecal excretion is not affected by level of Karanj in the diet. Feeding of expeller processed karanja cake at 24% of concentrate mixture caused adverse effect on N balance in lambs (Ravi et al., 2000). Even deoiled karanja cake feeding also caused reduction of N balance in growing calves (Konwar et al., 1987) and growing kids (Srivastava et al., 1990). Lower nitrogen intake was compensated by reduced urinary excretion due to increased demand of nitrogen and subsequent more recycling of urea across rumen epithelium (Kennedy and Milligan, 1980). It was further opined by these workers that mechanism by which this transfer occurs remain obscure. In contrary, Soren and Sastry (2009) observed positive Nbalance in lambs received differently processed karanj cake (water washing, lime treated or binder addition).
65.00 60.00 55.00 50.00 5
0
10
15
20
Level (%) of Karnaja in the diet
Kidney weight (g)
Linear (Kidney weight (g))
500.00 400.00 Gram
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300.00 y = -11.09x + 410.95 R² = 0.945
200.00 100.00 0.00 0
5
10
15
20
Leve (%) of Karanja in the diet Liver weight (g)
Linear (Liver weight (g))
Fig. 4. Regression analysis of hot carcass weight (kg) and weight of kidney and liver against percent of karanja cake in the diets.
aspartate aminotransferase (AST), alanine aminotransferase (ALT) (IU/l) and serum urea (mg/dl) were observed to be same in all groups indicating the residual ANF’s (Karanjin, pongamol, TIA and others) have not exerted any systemic effects in elevating or affecting any of the profiles studied. In contrast, Singh et al. (2006) observed higher activity of serum AST in lambs fed expeller and solvent extracted karanj cake. Earlier studies on unconventional oil cakes (Sal seed meal/Mahua seed cake) showed that the activity of serum transaminases increased in cow calves and sheep fed on concentrate mixtures containing such oil cake/meal for longer period (Garg et al., 1984; Singh et al., 1994). The values of serum protein observed in the present study are in normal reference values (Kaneko et al., 2008), however, albumin values were little higher than the reference range.
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4.5. Carcass characteristics, composition of Longissmus dorsi and organo-leptic evaluation of meat Similar to the study, hypertrophy of kidneys was observed by Singh et al. (2006) and Nagalakshmi et al. (2011) due to feeding of expeller and solvent extracted karanj cake. The reason attributed by them is due to effect of karanjin. Though, karanja did not elevate serum transaminases, it was good enough to cause hypertrophy of kidneys. It was already observed by feeding of detoxified karanja cake at higher levels affected the expression genes related to testicular function and testicular architecture (Dineshkumar et al., 2013). Similar to our results, protein and fat content in meat was not affected in lambs (Prabhu, 2002) and broilers (Panda et al., 2007) which were fed either raw or NaOHtreated solvent extracted karanj cake based diets. However, Soren et al. (2008) observed lower protein in the meat of lambs which were fed water-washed karanja seed cake compared to control lambs. 5. Conclusions The present study suggests that the detoxified karanja cake can be added as replacement of soybean meal (SBM) at low levels (more than 9 per cent of concentrate mixture). However, higher levels of replacement (above 9 per cent of concentrate mixture) warrant caution due to its adverse effect on growth performance, nutrient digestibility and important carcass trait like carcass weight. The effects are primarily caused due to residual ANF’s present in the cake which has affected the studied parameters. Conflict of interest We wish to confirm that there are no known conflicts of interest associated with this publication. Acknowledgments The authors thank Department of Biotechnology, for providing financial sanction for the project entitled “Detoxification and utilization of key agro forest based nonconventional oil cakes in the feeding of livestock” and The Director, NIANP Bangalore for providing necessary facilities. The authors further thank Dr. K. Ravikanth, Ayurvet Research Foundation, Baddi, Himachal Pradesh, India for analyzing samples for anti nutritional factors in the Karanj cake. We are also thankful for the technical support of Mr. H. S. Narayana Rao, Technician, NIANP for rendering assistance. References Alumot, E., Nistan, Z., 1961. The influence of soybean antitrypsin inhibitor on the intestine proteolysis of the chick. J. Nutr. 73, 71–77. AOAC, 2000. Official Methods of Analysis, Association of Official Analytical Chemists. 16th ed., Washington, DC. Bringi, N.V., Mukerjee, S.K., 1987. Karanja seed (Pongamiaglabra) oil. In: Bringi, N.V. (Ed.), Non-traditional Oilseeds and Oils in India. Oxford and IBH Publishing Co. Pvt. Ltd, New Delhi, pp. 143–166. Chitra, R., Sadasivam, S., 1986. A study of the trypsin inhibitor of black gram (Vigna mungo l hepper). Food Chem. 21, 315–320.
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Conway, E.J., 1957. Micro Diffusion Analysis and Volumetric Error, 4th ed. Cross by Lockwood and Sons, London. Dineshkumar, D., Rao, S.B.N., Jash, S., Elangovan, A.V., Hemalatha, S., 2011. Chemical composition and anti-nutritional factors in karanja (Pongamia pinnata) seed kernels and its in vitro evaluation. Indian J. Anim. Sci. 81, 478–483. Dineshkumar, D., Selvaraju, S., Parthipan, S., Thayakumar, A., Rajendran, D., Ravindra, J.P., Krishnamoorthy, P., Reddy, I.J., Rao, S.B.N., 2013. Effect of detoxified karanja (Pongamia spp.) cake on testicular architecture and semen production in ram lambs. Animal 7, 1697–1703. Garg, A.K., Agrawal, D.K., Nath, K., 1984. Effect of sal (Shorea robusta) seed meal tannins on serum enzymes, nutrient utilization and growth in growing calves. Agric. Wastes 11, 307–317. Gracey, J.F., 1981. Thornton’s Meat Hygiene, 7th ed. ELBS, Bailliere Tindall, East Sussex, UK, p. 111. Harvey, W.R., 1975. Least Square Analysis. United States Department of Agricultural Research Services (ARS), Washington, DC, USA. ICAR, 1998. Nutrient Requirement of Livestock and Poultry. Indian Council of Agricultural Research, New Delhi, India, p. 6. Kaneko, J.J., Harvey, J.W., Bruss, M.L., 2008. Appendix VIII blood analyte reference values in large animals. In: Clinical Biochemistry of Domestic Animals, 6th ed. Academic Press Elsevier, pp. 882–895. Kennedy, P.M., Milligan, L.P., 1980. The degradation and utilization of endogenous urea in the gastrointestinal tract of ruminants: a review. Can. J. Anim. Sci. 60, 205–221. Konwar, B.K., Banerjee, G.C., Mandal, L., 1987. Effect of feeding de-oiled karanja (Pongamiaglabra) cake on growing calves. Indian Vet. J. 64, 399–402. Makkar, H.P.S., Singh, B., Negi, S.S., 1990. Tannin levels and their degree of polymerization and specific activity in some agro-industrial byproducts. Biol. Wastes 31, 137–144. Mandal, 1985. Protease inhibitors and in vitro protein digestibility of defatted seed cakes of akashmoni and karanja. J. Am. Oil Chem. Soc. 62, 1124–1126. Nagalakshmi, D., Dhanalakshmi, K., Himabindu, D., 2011. Replacement of groundnut cake with sunflower and karanj seed cakes on performance, nutrient utilisation, immune response and carcass characteristics in Nellore lambs. Small Rumin. Res. 97, 12–20. Panda, A.K., Sastry, V.R.B., Kumar, A., Saha, S.K., 2006. Quantification of karanjin, tannin and trypsin inhibitors in raw and detoxified expeller and solvent extracted Karanj (Pongamia glabra) cake. Asian-Aust. J. Anim. Sci. 19, 1776–1783. Panda, A.K., Sastry, V.R.B., Mandal, A.B., Saha, S.K., Kumar, A., 2004. Effect of dietary incorporation of raw and processed karanj cake (Pongamia glabra) on nutrient retention and immune competence of broilerchicks. Anim. Nutr. Feed Technol. 4, 121–134. Panda, A.K., Sastry, V.R.B., Mandal, A.B., Saha, S.K., Kumar, A., 2007. Physico-chemical carcass characteristics and sensory attributes of meat of broiler chickens fed diets incorporated with processed karanj cake. Indian J. Poult. Sci. 42, 257–263. Prabhu, T.M., 2002. Clinico nutritional studies in lambs fed raw and detoxifiedkaranj (P. glabra vent) meal as protein supplement. PhD Thesis. Indian Veterinary Research Institute, Izatnagar, India. Prabhu, T.M., Devakumar, C., Sastry, V.R.B., Agrawal, D.K., 2002. Quantification of karanjin using high performance liquid chromatography in raw and detoxified karanj (Pongamia glabra vent) seed cake. Asian-Aust. J. Anim. Sci. 15, 416–420. Rangasamy, S., Seshadri, T.R., 1940. Pongamol: a new crystalline compound from pomgamia oil. Curr. Sci. 9, 179. Rao, S.B.N., Dineshkumar, D., Raju, P., Chandrasekaraiah, M., Thulasi, A., Lyjujose, V., Selvaraju, S., 2012. Effect of three energy levels on body weight change, nutrient utilization, rumen biochemical profiles and semen quality in rams. Indian J. Anim. Sci. 82, 108–113. Ravi, U., Singh, P., Garg, A.K., Agrawal, D.K., 2000. Performance of lambs fed expeller pressed and solvent extracted karanj (Pongamia pinnata) oil cake. Anim. Feed Sci. Technol. 88, 121–128. Ravikanth, K., Thakur, M., Singh, B., Saxena, M., 2009. TLC based method for standardization of Pongamia pinnata (Karanj) using karanjin as marker. Chromatographia 69, 597–599. Roy, D., Sharma, N.N., Khanna, R.N., 1977. Structure and synthesis of isoponga−avone: a new component of seeds of pongamia glabra. Indian J. Chem. 15B, 1138–1139. Singh, P., Agarwala, O.N., Charan, K., 1994. Blood values of lambs fed on deoiled Mahua (Bassia latifolia) seed cake. In: Somvanshi, R., Lokeshwar, R.R. (Eds.), Current Advances in Veterinary Sciences and Animal Production in India. , 1st ed. International Book Distributing Co., Lucknow, pp. 248–256. Singh, P., Sastry, V.R.B., Garg, A.K., Sharma, A.K., Singh, G.R., Agrawal, D.K., 2006. Effect of long term feeding of expeller pressed and solvent
Please cite this article in press as: Rao, S.B.N., Kumar, D.D., Effect of substitution of soybean meal by detoxified karanja cake on diet digestibility, growth, carcass and meat traits of sheep. Small Ruminant Res. (2015), http://dx.doi.org/10.1016/j.smallrumres.2015.03.010
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extracted karanj (Pongamia pinnata) seed cake on the performance of lambs. Anim. Feed Sci. Technol. 126, 157–167. Snedecor, G.W., Cochran, W.G., 1994. Statistical Methods, 8th ed. Oxford and IBH Pub. Co., Kolkata. Soren, N.M., Sastry, V.R.B., 2009. Replacement of soybean meal with processed karanj (Pongamia glabra) cake on the balances of karanjin and nutrients, as well as microbial protein synthesis in growing lamb. Anim. Feed Sci. Technol. 149, 16–29. Soren, N.M., Sastry, V.R.B., Saha, S.K., Kumar, A., 2007. Quantification of karanjin in raw and variously processed expeller pressed karanj (Pongamia glabra) cake using high performance liquid chromatography. Anim. Nutr. Feed Technol. 7, 261–268. Soren, N.M., Sastry, V.R.B., Saha, S.K., Mendiratta, S.K., 2008. Effect of feeding processed karanj (Pongamia glabra) cake on carcass characteristics
and meat sensory attributes of fattening lambs. Indian J. Anim. Sci. 78, 858–862. Srivastava, J.P., Gupta, B.S., Thakur, S., Verma, A.K., 1990. Utilization of deoiled karanj (Pongamia glabra) cake in kid grower rations. Indian J. Anim. Nutr. 7, 15–20. Vansoest, P.J., Robertson, J.B., Lewis, B.A., 1991. Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J. Dairy Sci. 74, 3583–3597. Vinay, B.J., Kanya, T.C.S., 2008. Effect of detoxification on the functional and nutritional quality of proteins of karanja seed meal. Food Chem. 106, 77–84. Wilson, P.A., Melmed, R.N., Hampe, M.V., Holt, S.J., 1978. Immunochemical study of interaction of soybean trypsin inhibitor with rat intestinal mucosa. Gut 19, 260L 265.
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Effect of substitution of soybean meal by detoxified karanja cake on diet digestibility, growth, carcass and meat traits of sheep S.B.N. Rao a,∗ , D. Dinesh Kumar b,1 a
Animal Nutrition Division, National Institute of Animal Nutrition and Physiology, Adugodi, Bangalore 560 030, India CNPq-TWAS Research Scholar, Laboratory of Animal Nutrition, Center for nuclear energy in Agriculture (CENA), University of Sao Paulo, Brazil b
a r t i c l e
i n f o
Article history: Received 17 July 2013 Received in revised form 12 March 2015 Accepted 12 March 2015 Available online xxx Keywords: Biochemical metabolites Carcass traits Digestibility Karanja cake Sheep
a b s t r a c t The non-conventional karanja cake is rich in protein (around 30% CP) and can be used in livestock feed as a protein source instead of conventional protein supplement cake like soybean meal (SBM), groundnut cake, etc. The present study was carried out to research the effect of partially substituted soybean meal with detoxified karanja cake (dKC) on performance of ram lambs. Twenty-four ram lambs were randomly divided into four groups (n = 6) and fed different levels (%) of detoxified karanja cake (0% replacement, control; 25% replacement, dKC-25; 50% replacement, dKC-50 and 75% replacement, dKC-75) in concentrate mixtures for 140 days. dKC was incorporated in the concentrate mixtures at the expense of soybean meal, maize grain and wheat bran at 9, 18 and 29% in dKC-25, dKC-50 and dKC-75, respectively on fresh basis. As the level of karanja in the diet increased, DMI was found to be decreasing significantly. Similar to these effects, N-retention was reduced leading to significant reduction of body weight in high karanja cake replaced groups. Similar trend was observed in OM, CP, and ADF digestilities and reason attributed to increased dietary level of karanja cake. However, detrimental effects were not observed on the levels of total protein, albumin, globulin, serum urea, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) indicating residual ANF present in dKC did not exert any adverse effects. The effects on hot carcass weight, weights of liver and testes are following a decreasing trend while that weight of kidney is increasing with level of karanja in the diet. Our findings highlights that the detoxified karanja cake can be added as replacement of soybean meal (SBM) at low levels. However, higher levels of replacement (above 9 per cent of concentrate mixture) warrant caution due to its adverse effect on studied parameters. © 2015 Elsevier B.V. All rights reserved.
1. Introduction Karanja (Pongamia spp.) is grown in parts of humid tropical regions of Asia and Oceania. The seed contains 20–30%
∗ Corresponding author. Tel.: +91 8025711304; fax: +91 8025711420. E-mail addresses: [email protected], [email protected] (S.B.N. Rao). 1 The second author is at present pursuing his Ph.D. in Brazil. The reported work was conducted in Bangalore. He worked as Research Fellow in the project.
protein, 27–39% oil and a group of furano-flavonoids that constitutes 5–6% by weight of the oil (Bringi and Mukerjee, 1987). In a recent study, large variations were observed in seed size, composition in terms of protein and fat, incriminating factors like karanjin, pongamol and trypsin inhibitor activity (TIA) collected from different regions of Karnataka (Dineshkumar et al., 2011). The seed kernel after extraction of oil is rich in protein (around 30% CP) which can be used in animal and poultry feed as a protein source (Konwar et al., 1987; Vinay and Kanya, 2008). However, raw expeller karanja cake (rKC) is not commonly used as
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a feed for livestock due to its poor palatability and presence of antinutritional factors (ANFs), i.e. furanoflavones like karanjin, pongamol and certain other polyphenolic compounds (Bringi and Mukerjee, 1987; Prabhu et al., 2002) leading to depression in performance of livestock (Srivastava et al., 1990; Nagalakshmi et al., 2011; Panda et al., 2004). Various methods, namely refluxing with 2% HCl (Mandal, 1985) water leaching, solvent extraction, acid and alkali treatment, autoclaving were adopted to improve palatability and nutritive value of detoxify karanja cake. Among all the methods, solvent extraction was found to be more efficient to remove karanjin (Prabhu et al., 2002) and is being commonly used as a method to detoxify karanja cake. Singh et al. (2006) reported that long term feeding of expeller karanja cake or solvent extracted karanja cake had deleterious effects on the nutrient utilization, blood biochemical profile, rumen fermentation pattern, carcass characteristics and manifested clinico-pathological changes in bone and tissues of vital organs. Further studies indicated that inclusion of expeller karanja cake at 12% in complete diets constituting 390 mg karanjin per kg depressed the performance in lambs (Nagalakshmi et al., 2011). Keeping in view the adverse effects exerted by feeding of karanja cake, present study aimed to fill the gaps in scale up of detoxification methodology of karanja cake at industrial scale and its possible utilization in livestock rations replacing soybean cake. The effects on physiological responses in terms of gene expression of LH receptor and IGF-I genes related to testicular function, testicular architecture and hormones was already reported (Dineshkumar et al., 2013). The objective of the particular experiment is to test different levels of detoxified karanja cake in rations of growing lambs based on its effects on intake, digestibility, growth, carcass and meat traits. 2. Materials and methods 2.1. Source of detoxified karanja seed cake (dKC) Bulk production of detoxified meal required for feeding trial was collected from industrial partners (M/s Ayurvet Pvt. Limited, Baddi, Himachal Pradesh, India and M/s Ganesh Scientific Research Foundation, New Delhi,
India). A process was developed to detoxify karanja seeds initially by crushing and removing the oil using hexane. Then the meal was further detoxified by soaking the seeds in aqueous methanol mixture and drying. 2.2. Feed preparation The requirements of crude protein (CP) and total digestible nutrients (TDN) suggested by Indian Council of Agricultural Research (ICAR, 1998) formed the guidelines for feeding lambs. The required quantities of feed ingredients like finger millet (Eleusine corocana) straw, maize (Zea mays) grain, soybean (Glycine max) meal, wheat (Triticum aestivum) bran, mineral mixture and salt were procured from commercial source for the entire duration of the experiment Maize grain and soybean meal were ground in hammer mill and mixed with wheat bran, mineral-vitamin premix and salt in horizontal mixer. Four iso-nitrogenous and iso-caloric concentrate mixtures were prepared by making minor adjustments in the proportions of wheat bran and maize grain were made in the formulation (Table 1). 2.3. Experimental animals and housing 24 ram lambs were used in this experiment (body weight: 13.7 ± 0.5 kg; aged around 6 months) for a period of 140 days in a permanent open type shed having provision for tying the animals in separate enclosures and also feeding trough. Ram lambs were fed individually and reared under hygienic and uniform managerial conditions throughout the experiment. Ram lambs were offered clean water 2–3 times daily for the entire duration. They were dewormed using broad spectrum anthelmentic (albendazole®) at the rate of 10 mg per kg body weight twice at 21 days interval at the beginning of the trial and all the animals were confirmed to be parasite free by faecal examination. Ram lambs were randomly divided into four groups (n = 6) and were offered concentrate mixture incorporating dKC as a replacement of SBM. Different levels (%) of karanja cake were fed (0% replacement, control; 25% replacement—9% dKC in concentrate mixture on fresh basis, dKC-25; 50% replacement—18% dKC in concentrate mixture on fresh basis, dKC-50 and 75% replacement—29% dKC in concentrate mixture on fresh basis, dKC-75). dKC was incorporated in concentrate mixtures (on fresh basis) at 9, 18 and 29% in dKC-25, dKC-50 and dKC-75, respectively. The lambs were offered the respective concentrate mixtures daily between 9:00 and 9:30 h, to meet protein requirements (ICAR, 1998) for maintenance and expected daily gains of 75 g. The lambs were fed concentrate mixture around 1.6–1.7 percent of bodyweight. The remaining requirements were met through feeding of chaffed finger millet (Eleusince corocana) straw (∼1.6% of body weight), which was offered after 2:00 h of feeding concentrate mixture fed to ensure its maximum consumption. On an average lambs consumed around 250–350 g each of concentrate and ragi straw across different treatment groups. All lambs were weighed individually at weekly intervals to assess body weight changes and growth
Table 1 Ingredients and chemical composition of feeds used in the experiments. Item
dKC 0
dKC 25
dKC 50
dKC 75
Ragistraw
Ingredients (%) Maize grain Soybean meal Karanja detoxified cake Wheat bran Mineral mixtureA Salt Dry matter (%) Organic matter (% DM) Ether extract (% DM) Crude protein (% DM) Acid detergent fibre (% DM) Karanjin (%)a Trypsin inhibitor activity (g/g)a
31 30 0 36 2 1 96.84 91.70 2.61 22.75 8.89 – –
31 22.5 9 34.5 2 1 97.26 92.13 3.00 22.57 8.63 0.0027 35.1
33 15 18 31 2 1 97.51 93.28 3.73 22.30 9.05 0.0054 70.2
34 7.5 29 26.5 2 1 97.01 93.69 3.35 21.72 8.57 0.0087 113.1
93.53 91.58 1.18 4.05 43.35 – –
dKC-0, detoxified karanja cake 0% replacement with Soybean cake; dKC-50, detoxified karanja cake 50% replacement with Soybean cake and dKC-75, detoxified karanja cake 75% replacement with Soybean cake. A Commercial mineral mixture with the composition per kg: Cobalt-150 mg, copper-1200 mg, iodine-325 mg, iron-5000 mg, magnesium-6000 mg, potassium-1500 mg, selenium-100 mg, sodium-10 mg, sulphur-5.9 mg, zinc-0.942%, DL Methinonine-9600 mg, L-lysine, mono hydrochloride-4400 mg, calcium-24%, phosphorous-12% was used. a Calculated values on the basis of concentrations of detoxified karnaja cake.
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2.4. Metabolism trial After 90 days of the experimental feeding, a 6-days metabolism trial was carried out on all lambs (n = 24) involving total faecal and urine collection. This collection is preceded by 2 days adaptation period for lambs’ acclimatization to faecal and urine bags. During the trial period, all the lambs were tied with cotton ropes. The faeces collection was facilitated by tying tarpaulin faecal bags and urine was collected in polythene bags. The contents of faeces and urine from bags were cleared at regular intervals manually in the airtight plastic containers contained known quantity of H2 SO4 . Daily feed offered, residues left and faeces and urine excreted was recorded on 24-h basis. Feeds, refusals and an aliquot of faeces were dried at 100 ± 5 ◦ C for 24 h for the determination of DM. The dry samples were ground to pass through a 1-mm sieve and pooled for proximate analysis. Day to day fresh samples of faeces (1/10) was preserved with 1:4 sulphuric acid for N estimation. 2.5. Collection and analysis of rumen liquor and blood samples After the metabolism trial, rumen liquor was collected from all the sheep with the help of stomach tube. Little suction was applied to collect the rumen liquor from the sheep (Rao et al., 2012). The pH of rumen liquor was recorded immediately after collection using a pre-calibrated digital pH Meter. The strained rumen liquor was analyzed for ammoniaN (Conway, 1957), total-N and trichloroacetic acid precipitable-N (TCA ppt.-N) (AOAC, 2000). Blood was collected from jugular vein. Serum was separated and stored at −80◦ C till analyzed. Biochemical profiles such serum protein (g/dl), albumin (g/dl), glucose (g/dl), serum urea (mg/dl) and enzymes such as alanine amino tranferease (ALT) and aspartate aminotransferase (AST) (IU/l) were estimated using biochemical kits purchased from Span Diagnostics Limited, Surat, Gujarat, India. The globulin concentration was measured as difference between total protein and albumin. 2.6. Laboratory analysis Proximate analysis of the concentrate mixtures, finger millet straw and faeces viz. CP, EE and ash were determined by the methods of AOAC (2000). Acid detergent fibre (ADF) was estimated using the procedure of Vansoest et al. (1991). The karanjin was estimated by separation of components by TLC on pre-coated silica gel G 60 F254 plates developed on toluene: ethyl acetate (7:3 v/v) and detection at 260 nm in absorbance mode (Ravikanth et al., 2009) and Trypsin inhibitory activity (TIU/g) indirectly by inhibiting the activity of trypsin. A synthetic substrate Nu-benzoyl OL-arginine p-nitroanilide (BAPNA) is subjected to hydrolysis by trypsin to produce yellow coloured p-nitroanilide. The degree of inhibition by the extract of the yellow colour production is measured at 410 nm (Chitra and Sadasivam, 1986). 2.7. Slaughter and carcass evaluation Feed was withheld overnight with free access to water and animals were slaughtered in a local abattoir by Halal method (Gracey, 1981). After slaughter, the head and feet were removed and the animals were partially skinned lying on their back on the floor. Thereafter, the animals were suspended by the hind legs for further skinning. Carcass and noncarcass components were weighed immediately after slaughter. Lungs, trachea and heart were weighed as one piece and designated as pluck. Non-carcass components included head, skin, feet, digestive tract, liver, spleen, pancreas and pluck. Weight of digestive contents was computed as the difference between full and empty digestive tract. The empty live weight was computed as the difference between slaughter weight and weight of digesta content. 2.8. Meat quality evaluation The longissimus dorsi muscle was collected, within 20 min of slaughter, trimmed of fat and frozen at −80 ◦ C till analyzed for meat quality traits. The pH of minced L. dorsi muscle was recorded by using a digital pH meter. Proximate composition of longissimus dorsi was determined on muscle
800.00 Dry maer intake (g/day)
rate. The individual intake of nutrients such as DM, CP, DCP and TDN per unit gain was calculated by dividing intake of nutrients obtained from individual animals during sub trial for arriving values of nutrient conversion efficiency.
3
700.00 600.00 500.00 400.00 300.00 200.00 y = -12.266x + 688.62 R² = 0.985
100.00 0.00 0
2
4
6
8
10
12
14
16
Level (%) of Karanja cake in the diet DMI (g/day)
Linear (DMI (g/day))
Fig. 1. Regression analysis of average dry matter intake (Y-axis) and percent of Karanja cake in the diets.
samples according to AOAC (2000). The sensory evaluation of pressurecooked meat samples for colour, odour, juiciness, tenderness and overall palatability was performed by a taste panel consisting of 10 panelists using 9 point scale (9 = like extremely, 1 = dislike extremely).
2.9. Statistical analysis Individual sheep data was considered as a replicate and a one-way analysis of variance (ANOVA) was done using completely randomized design (Snedecor and Cochran, 1994) by a mathematical model of Harvey (1975). SPSS is a software package used for statistical analysis (SPSS Inc., Chicago, IL, USA) for analysis of variance assuming for independent constant variance structure with post-hoc tukey to find the pair wise significance between treatments. A P-value of less than or equal to 0.05 was accepted to indicate statistical significance. Repeated measure analysis was performed on for body weights recorded at different periods (weeks) using JMP® 11.2.0 software by fitting a model with a role variable as body weights, constructing model effects (treatment and period) and interactions (Treatment × Period) and generated least square means were subjected for significance using tukey post-hoc. Linear regression analysis was performed on different parameters such as dry matter intake, N excretion (faeces) and urine, N retained, hot carcass weight, organ weighs (y variable) with % of Karanja in the diet to construct a linear bivariate fit.
3. Results 3.1. Chemical composition of concentrate mixtures CP content (%) of concentrate mixtures was found to be 21.7–22.8 in all groups (Table 1). EE% was found to be ranging from 2.6 in control to 3.7 in dKC-50. Increased EE in case of dKC groups could be due to more EE content of dKC used in the experiment (3.85%). dKC had shown residual concentration of Karanjin, 0.03% W/W and TIA, 390 g/g. 3.2. Intake, digestibility and balance of nitrogen The daily dry matter intake of lambs was regressed against per cent of incorporation of Karanja in the diet indicated significantly negative correlation (Fig. 1) indicating as the level of karanja increased in the diet the DMI is following a decreased trend. This was also reflected on reduced DCP intake (g/d) in dKC-50 and dKC-75 groups compared to control and dKC-25. Compared to control, dKC-50 and dKC-75 groups had low digestibilities of DM, OM, ADF and T-CHO. CP digestibility was lowest in dKC-75 (Table 2).
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4 Table 2 Intake, digestibility in different groups. Attributes
Diet
SEM
dKC 0
dKC 25
dKC 50
dKC 75
Dry matter intake (g/d) DOM intake (g/d) DCP intake (g/d) TDN intake (g/d)
684.81 437.04 73.02b 436.50a
643.92 407.84 69.36b 411.49a,b
565.16 346.63 57.28a,b 350.71a,b
510.75 317.13 46.17a 315.92b
Nutrient digestibility (%) Organic matter Crude protein ADF
69.71b 79.29b 47.66b
68.97a,b 79.79b 44.62b
66.25a 75.68a,b 37.57a
26.09 17.49 3.25 17.31
66.68a,b 68.97a 37.61a
0.49 1.79 1.31
dKC-0, detoxified karanja cake 0% replacement with Soybean cake; dKC-50, detoxified karanja cake 50% replacement with Soybean cake and dKC-75, detoxified karanja cake 75% replacement with Soybean cake. Means bearing different superscripts (a and b) in a row differ significantly (P < 0.05).
groups (Table 3). Differences in total protein, albumin, aspartate aminotransferase (AST), alanine aminotransferase (ALT) (IU/l) and serum urea (mg/dl) were observed to be same in all groups indicating the residual ANF’s (Karanjin, pongamol, TIA and others) have not exerted any systemic effects in elevating or affecting any of the profiles studied. 3.5. Carcass characteristics, composition of Longissmus dorsi and organo-leptic evaluation of meat Fig. 2. Bodyweight least square means (kg) across different treatments (1, dKC-0, detoxified karanja cake 0% replacement with Soybean cake 19.34; 2, dKC-25, detoxified karanja cake, 18.64; 3, dKC, 50, 50% replacement with Soybean cake, 17.02; and 4, dKC-75, detoxified karanja cake 75% replacement with Soybean cake 15.52 kg).
3.3. Body weights during experimental period
Hotcarcass weight was following declining trend as level of karanja cake increased in diet (Fig. 3). Similar to hot carcass weight, liver, heart and testes weights following declining trend. However, weight of kidney was following a increasing trend. Analyzed parameters like dry matter, fat, protein and ash were found to be similar in all treatment groups indicating no adverse effect of feeding dKC on chemical composition of meat. Organoleptic evaluation in terms of like appearance, colour, texture and consistency, aroma and flavor and overall acceptability were found to be same in all tested groups (Table 4).
The least square means of body weights across different treatments presented (Fig. 2). The average body weights recorded across different treatments were similar only up to 25% of replacement. At higher levels of replacements (50 and 75%), body weights were significantly (P < 0.05) reduced.
4. Discussion
3.4. Rumen metabolites and serum biochemical profiles
4.1. Chemical composition of concentrate mixtures
Rumen fermentation profiles such as pH, ammonia-N, TCA-ppt N, total-N were similar (P > 0.05) in all treatment
Karanjin contents of expeller/solvent extracted Karanj seed cake reported by various workers 0.32% by
Table 3 Rumen metabolites, serum biochemical profiles of lambs. Attributes
pH Ammonia nitrogen (mg/100 ml SRL) Total nitrogen (mg/100 ml SRL) TCA ppt nitrogen (mg/100 ml SRL) Serum protein (g/100 ml) Serum albumin (g/100 ml) Urea (mg/dl) Aspartate aminotransferase (IU/l) Alanine aminotranferease (IU/l)
Diet
SEM
dKC 0
dKC 25
dKC 50
dKC 75
6.46 40.61 101.62 47.37 7.00 4.83 60.10 143.6 38.61
6.44 45.29 100.28 50.96 6.93 4.91 58.70 140.6 34.10
6.42 35.93 99.17 56.66 6.97 4.89 50.45 147.8 36.86
6.40 33.88 99.54 60.48 6.40 4.48 56.24 145.8 30.97
13.27 2.29 2.67 0.30 0.57 0.11 1.95 9.51 3.16
dKC-0, detoxified karanja cake 0% replacement with Soybean cake; dKC-50, detoxified karanja cake 50% replacement with Soybean cake and dKC-75, detoxified karanja cake 75% replacement with Soybean cake. Means bearing different superscripts (a and b) in a row differ significantly (P < 0.05).
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5
Table 4 Caracass characteristics, composition of Longissmus dorsi and organo-leptic evaluation of meat by semi-trained panelists. Attributes
Diet
SEM
dKC-0
dKC-25
dKC-50
dKC-75
Live body weight (kg) after 12 h of fasting Hot carcass weight (kg)
24.08b 11.04b
24.75b 11.10b
22.33a,b 9.84a,b
17.75a 8.05a
0.93 0.43
Weight of vital organs (g) Liver Kidneys Heart Testes Spleen
391b 57 122 172 51
384b 60 144 250 57
325a,b 61 140 202 59
271a 68 107 164 48
16 3 6 33 3
Chemical composition (% on fresh basis) Dry matter (%) Total fat (%) Ash (%) Protein (%) pH
27.30 5.22 1.01 19.82 5.65a
28.32 5.28 1.06 21.17 5.73b
25.91 3.61 1.10 20.47 5.66a
27.63 4.16 1.15 21.93 6.05c
0.45 0.28 0.02 0.35 0.04
Organoleptic evaluation Appearance Colour Texture and consistency Aroma and flavour Overall acceptability
6.63 6.63 5.50 5.94 6.38
6.13 6.13 6.13 5.88 6.38
5.63 5.63 6.38 5.94 6.50
– – – – –
0.27 0.27 0.26 0.21 0.26
g/day
dKC-0, detoxified karanja cake 0% replacement with Soybean cake; dKC-50, detoxified karanja cake 50% replacement with Soybean cake and dKC-75, detoxified karanja cake 75% replacement with Soybean cake. Means bearing different superscripts (a,b,c) in a row differ significantly (P < 0.05).
4.50 4.00 3.50 3.00 2.50 2.00 1.50 1.00 0.50 0.00
to this, Makkar et al. (1990) have reported lower total tannins and condensed tannins content in karanj cake and protein precipitation was not reported. The calculated concentrations of karanjin and TIA in the present study have increased linearly as level dKC increased in the concentrate mixtures (0.0027–0.0087 for karanjin and 35.1 and 113.1 for TIA).
y = -0.0802x + 3.9291 R² = 0.8527
0
2
4
6
8
10
12
14
16
Level (%) of Karanja cake in the diet
g/day
N- excreon (Urine)
Linear (N- excreon (Urine))
9.00 8.00 7.00 6.00 5.00 4.00 3.00 2.00 1.00 0.00
y = -0.2241x + 8.075 R² = 0.9818 0
2
4
6
8
10
12
14
16
Level (%) of Karanja in the diet N-retenon
Linear (N-retenon )
Fig. 3. Regression analysis of N-excretion (Urine) and N-retained (Y-axis) against percent of Karanja cake in the diets.
Nagalakshmi et al. (2011), 0.32% by Panda et al. (2006), 0.19% by Prabhu et al. (2002) and 0.29% by Soren et al. (2007). Vinay and Kanya (2008) have reported 0.94% of tannins along with other anti-nutritional factors, such as phytate (0.65%) and trypsin inhibitors (39 IU/mg). Contrary
4.2. Intake, digestibility and balance of nitrogen Many studies involved with feeding of either solvent extracted karanja cake or expeller karanja cake were reported (Konwar et al., 1987 in crossbred bull calves; Srivastava et al., 1990 in growing kids; Singh et al., 2006; Ravi et al., 2000; Prabhu, 2002; Soren and Sastry, 2009; Nagalakshmi et al., 2011 in growing lambs). As observed in present study reduction in feed intake was observed in cross-bred bulls fed 26.5% deoiled karanja cake based concentrate mixture (Konwar et al., 1987) and in lambs fed with concentrate mixtures containing either 24% expeller karanja cake or 20% deoiled karanja cake (Singh et al., 2006) or 22.5% processed deoiled karanja cake (Soren and Sastry, 2009) or 12% expeller karanja cake (Nagalakshmi et al., 2011). On the contrary, no effect on DMI has been reported in goat kids receiving deoiled karanja cake at 40% level in concentrate mixture (Srivastava et al., 1990). Other workers (Ravi et al., 2000; Prabhu, 2002) also reported similar observations on dry matter intake in growing lambs receiving de-oiled karanja cake at 20% level. Similar to our results, reduction in nutrient digestibility was observed (Srivastava et al., 1990) in goat kids receiving 12% de-oiled karanja cake and 24% expeller karanja cake in lambs by Ravi et al. (2000). However, no effect on DM, OM digestibilities (Soren
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4.3. Body weights during experimental period The results obtained in the present experiment are consistent with earlier similar experiments involved with feeding karanja cake (Ravi et al., 2000; Soren et al., 2008; Nagalakshmi et al., 2011) in growing lambs fed either expeller or processed karanja cake. 4.4. Rumen metabolites and serum biochemical profiles In line with the current study, Singh et al. (2006) found no significant differences in pH, NH3 -N, total-N and TCA ppt.-N in the rumen liquor of lambs fed expeller pressed (24.0%) and solvent extracted karanj cake (20.0%) for a period of 255 days. Differences in total protein, albumin,
14.00 12.00
kg
10.00 8.00 6.00 4.00
y = -0.2121x + 11.519 R² = 0.8822
2.00 0.00
5
0
10
15
20
Level (%) of Karanja in the diet Hot carcass weight (Kg)
Linear (Hot carcass weight (Kg))
80.00 75.00 y = 0.3696x + 58.414 R² = 0.5624
70.00 Gram
and Sastry, 2009) and nutrient retention (Panda et al., 2004) was observed with incorporation of raw or processed expeller or solvent extracted karanja cake in lambs and broilers, respectively. The negative effects of karanja cake feeding on intake, digestibility and subsequent nutritive value of diet mainly because of residual ANFs like karanjin, trypsin inhibitors, pongamol, pongapin and lanceolatin B present in residual oil portion of karanja cake which might have affected the digestibility of these nutrients because karanjin possesses the insecticidal and antibacterial properties and also phenolic compounds which are well known for depressing the nutrients digestibility (Rangasamy and Seshadri, 1940; Roy et al., 1977). The reduction in protein digestibility observed the present study might be due to residual tryspin inhibitors and tannins present in karanja cake. The adverse effect of trypsin inhibitors is mainly on the pancreas, which responds to the inhibitors by enhanced synthesis and secretion of proteolytic enzymes. The pancreatic enzyme secretion is regulated by a negative feedback mechanism mediated by intestinal trypsin and chymotrypsin, and complex formation of trypsin with the inhibitors leading to a reduction of free trypsin in the small intestine (Alumot and Nistan, 1961). This reduction activates the pancreas-stimulating hormone, cholecystokinin, the release of which from the intestinal mucosa is inhibited by free trypsin (Wilson et al., 1978). Regression analysis of N-retained (g/day) with % dietary incorporation of dKC indicated amount of N-retained is linearly decreased as the level of dKC increased in the diet. This is primarily due to low intake and subsequent urinary excretion of nitrogen (Fig. 4). However, faecal excretion is not affected by level of Karanj in the diet. Feeding of expeller processed karanja cake at 24% of concentrate mixture caused adverse effect on N balance in lambs (Ravi et al., 2000). Even deoiled karanja cake feeding also caused reduction of N balance in growing calves (Konwar et al., 1987) and growing kids (Srivastava et al., 1990). Lower nitrogen intake was compensated by reduced urinary excretion due to increased demand of nitrogen and subsequent more recycling of urea across rumen epithelium (Kennedy and Milligan, 1980). It was further opined by these workers that mechanism by which this transfer occurs remain obscure. In contrary, Soren and Sastry (2009) observed positive Nbalance in lambs received differently processed karanj cake (water washing, lime treated or binder addition).
65.00 60.00 55.00 50.00 5
0
10
15
20
Level (%) of Karnaja in the diet
Kidney weight (g)
Linear (Kidney weight (g))
500.00 400.00 Gram
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300.00 y = -11.09x + 410.95 R² = 0.945
200.00 100.00 0.00 0
5
10
15
20
Leve (%) of Karanja in the diet Liver weight (g)
Linear (Liver weight (g))
Fig. 4. Regression analysis of hot carcass weight (kg) and weight of kidney and liver against percent of karanja cake in the diets.
aspartate aminotransferase (AST), alanine aminotransferase (ALT) (IU/l) and serum urea (mg/dl) were observed to be same in all groups indicating the residual ANF’s (Karanjin, pongamol, TIA and others) have not exerted any systemic effects in elevating or affecting any of the profiles studied. In contrast, Singh et al. (2006) observed higher activity of serum AST in lambs fed expeller and solvent extracted karanj cake. Earlier studies on unconventional oil cakes (Sal seed meal/Mahua seed cake) showed that the activity of serum transaminases increased in cow calves and sheep fed on concentrate mixtures containing such oil cake/meal for longer period (Garg et al., 1984; Singh et al., 1994). The values of serum protein observed in the present study are in normal reference values (Kaneko et al., 2008), however, albumin values were little higher than the reference range.
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4.5. Carcass characteristics, composition of Longissmus dorsi and organo-leptic evaluation of meat Similar to the study, hypertrophy of kidneys was observed by Singh et al. (2006) and Nagalakshmi et al. (2011) due to feeding of expeller and solvent extracted karanj cake. The reason attributed by them is due to effect of karanjin. Though, karanja did not elevate serum transaminases, it was good enough to cause hypertrophy of kidneys. It was already observed by feeding of detoxified karanja cake at higher levels affected the expression genes related to testicular function and testicular architecture (Dineshkumar et al., 2013). Similar to our results, protein and fat content in meat was not affected in lambs (Prabhu, 2002) and broilers (Panda et al., 2007) which were fed either raw or NaOHtreated solvent extracted karanj cake based diets. However, Soren et al. (2008) observed lower protein in the meat of lambs which were fed water-washed karanja seed cake compared to control lambs. 5. Conclusions The present study suggests that the detoxified karanja cake can be added as replacement of soybean meal (SBM) at low levels (more than 9 per cent of concentrate mixture). However, higher levels of replacement (above 9 per cent of concentrate mixture) warrant caution due to its adverse effect on growth performance, nutrient digestibility and important carcass trait like carcass weight. The effects are primarily caused due to residual ANF’s present in the cake which has affected the studied parameters. Conflict of interest We wish to confirm that there are no known conflicts of interest associated with this publication. Acknowledgments The authors thank Department of Biotechnology, for providing financial sanction for the project entitled “Detoxification and utilization of key agro forest based nonconventional oil cakes in the feeding of livestock” and The Director, NIANP Bangalore for providing necessary facilities. The authors further thank Dr. K. Ravikanth, Ayurvet Research Foundation, Baddi, Himachal Pradesh, India for analyzing samples for anti nutritional factors in the Karanj cake. We are also thankful for the technical support of Mr. H. S. Narayana Rao, Technician, NIANP for rendering assistance. References Alumot, E., Nistan, Z., 1961. The influence of soybean antitrypsin inhibitor on the intestine proteolysis of the chick. J. Nutr. 73, 71–77. AOAC, 2000. Official Methods of Analysis, Association of Official Analytical Chemists. 16th ed., Washington, DC. Bringi, N.V., Mukerjee, S.K., 1987. Karanja seed (Pongamiaglabra) oil. In: Bringi, N.V. (Ed.), Non-traditional Oilseeds and Oils in India. Oxford and IBH Publishing Co. Pvt. Ltd, New Delhi, pp. 143–166. Chitra, R., Sadasivam, S., 1986. A study of the trypsin inhibitor of black gram (Vigna mungo l hepper). Food Chem. 21, 315–320.
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Conway, E.J., 1957. Micro Diffusion Analysis and Volumetric Error, 4th ed. Cross by Lockwood and Sons, London. Dineshkumar, D., Rao, S.B.N., Jash, S., Elangovan, A.V., Hemalatha, S., 2011. Chemical composition and anti-nutritional factors in karanja (Pongamia pinnata) seed kernels and its in vitro evaluation. Indian J. Anim. Sci. 81, 478–483. Dineshkumar, D., Selvaraju, S., Parthipan, S., Thayakumar, A., Rajendran, D., Ravindra, J.P., Krishnamoorthy, P., Reddy, I.J., Rao, S.B.N., 2013. Effect of detoxified karanja (Pongamia spp.) cake on testicular architecture and semen production in ram lambs. Animal 7, 1697–1703. Garg, A.K., Agrawal, D.K., Nath, K., 1984. Effect of sal (Shorea robusta) seed meal tannins on serum enzymes, nutrient utilization and growth in growing calves. Agric. Wastes 11, 307–317. Gracey, J.F., 1981. Thornton’s Meat Hygiene, 7th ed. ELBS, Bailliere Tindall, East Sussex, UK, p. 111. Harvey, W.R., 1975. Least Square Analysis. United States Department of Agricultural Research Services (ARS), Washington, DC, USA. ICAR, 1998. Nutrient Requirement of Livestock and Poultry. Indian Council of Agricultural Research, New Delhi, India, p. 6. Kaneko, J.J., Harvey, J.W., Bruss, M.L., 2008. Appendix VIII blood analyte reference values in large animals. In: Clinical Biochemistry of Domestic Animals, 6th ed. Academic Press Elsevier, pp. 882–895. Kennedy, P.M., Milligan, L.P., 1980. The degradation and utilization of endogenous urea in the gastrointestinal tract of ruminants: a review. Can. J. Anim. Sci. 60, 205–221. Konwar, B.K., Banerjee, G.C., Mandal, L., 1987. Effect of feeding de-oiled karanja (Pongamiaglabra) cake on growing calves. Indian Vet. J. 64, 399–402. Makkar, H.P.S., Singh, B., Negi, S.S., 1990. Tannin levels and their degree of polymerization and specific activity in some agro-industrial byproducts. Biol. Wastes 31, 137–144. Mandal, 1985. Protease inhibitors and in vitro protein digestibility of defatted seed cakes of akashmoni and karanja. J. Am. Oil Chem. Soc. 62, 1124–1126. Nagalakshmi, D., Dhanalakshmi, K., Himabindu, D., 2011. Replacement of groundnut cake with sunflower and karanj seed cakes on performance, nutrient utilisation, immune response and carcass characteristics in Nellore lambs. Small Rumin. Res. 97, 12–20. Panda, A.K., Sastry, V.R.B., Kumar, A., Saha, S.K., 2006. Quantification of karanjin, tannin and trypsin inhibitors in raw and detoxified expeller and solvent extracted Karanj (Pongamia glabra) cake. Asian-Aust. J. Anim. Sci. 19, 1776–1783. Panda, A.K., Sastry, V.R.B., Mandal, A.B., Saha, S.K., Kumar, A., 2004. Effect of dietary incorporation of raw and processed karanj cake (Pongamia glabra) on nutrient retention and immune competence of broilerchicks. Anim. Nutr. Feed Technol. 4, 121–134. Panda, A.K., Sastry, V.R.B., Mandal, A.B., Saha, S.K., Kumar, A., 2007. Physico-chemical carcass characteristics and sensory attributes of meat of broiler chickens fed diets incorporated with processed karanj cake. Indian J. Poult. Sci. 42, 257–263. Prabhu, T.M., 2002. Clinico nutritional studies in lambs fed raw and detoxifiedkaranj (P. glabra vent) meal as protein supplement. PhD Thesis. Indian Veterinary Research Institute, Izatnagar, India. Prabhu, T.M., Devakumar, C., Sastry, V.R.B., Agrawal, D.K., 2002. Quantification of karanjin using high performance liquid chromatography in raw and detoxified karanj (Pongamia glabra vent) seed cake. Asian-Aust. J. Anim. Sci. 15, 416–420. Rangasamy, S., Seshadri, T.R., 1940. Pongamol: a new crystalline compound from pomgamia oil. Curr. Sci. 9, 179. Rao, S.B.N., Dineshkumar, D., Raju, P., Chandrasekaraiah, M., Thulasi, A., Lyjujose, V., Selvaraju, S., 2012. Effect of three energy levels on body weight change, nutrient utilization, rumen biochemical profiles and semen quality in rams. Indian J. Anim. Sci. 82, 108–113. Ravi, U., Singh, P., Garg, A.K., Agrawal, D.K., 2000. Performance of lambs fed expeller pressed and solvent extracted karanj (Pongamia pinnata) oil cake. Anim. Feed Sci. Technol. 88, 121–128. Ravikanth, K., Thakur, M., Singh, B., Saxena, M., 2009. TLC based method for standardization of Pongamia pinnata (Karanj) using karanjin as marker. Chromatographia 69, 597–599. Roy, D., Sharma, N.N., Khanna, R.N., 1977. Structure and synthesis of isoponga−avone: a new component of seeds of pongamia glabra. Indian J. Chem. 15B, 1138–1139. Singh, P., Agarwala, O.N., Charan, K., 1994. Blood values of lambs fed on deoiled Mahua (Bassia latifolia) seed cake. In: Somvanshi, R., Lokeshwar, R.R. (Eds.), Current Advances in Veterinary Sciences and Animal Production in India. , 1st ed. International Book Distributing Co., Lucknow, pp. 248–256. Singh, P., Sastry, V.R.B., Garg, A.K., Sharma, A.K., Singh, G.R., Agrawal, D.K., 2006. Effect of long term feeding of expeller pressed and solvent
Please cite this article in press as: Rao, S.B.N., Kumar, D.D., Effect of substitution of soybean meal by detoxified karanja cake on diet digestibility, growth, carcass and meat traits of sheep. Small Ruminant Res. (2015), http://dx.doi.org/10.1016/j.smallrumres.2015.03.010
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extracted karanj (Pongamia pinnata) seed cake on the performance of lambs. Anim. Feed Sci. Technol. 126, 157–167. Snedecor, G.W., Cochran, W.G., 1994. Statistical Methods, 8th ed. Oxford and IBH Pub. Co., Kolkata. Soren, N.M., Sastry, V.R.B., 2009. Replacement of soybean meal with processed karanj (Pongamia glabra) cake on the balances of karanjin and nutrients, as well as microbial protein synthesis in growing lamb. Anim. Feed Sci. Technol. 149, 16–29. Soren, N.M., Sastry, V.R.B., Saha, S.K., Kumar, A., 2007. Quantification of karanjin in raw and variously processed expeller pressed karanj (Pongamia glabra) cake using high performance liquid chromatography. Anim. Nutr. Feed Technol. 7, 261–268. Soren, N.M., Sastry, V.R.B., Saha, S.K., Mendiratta, S.K., 2008. Effect of feeding processed karanj (Pongamia glabra) cake on carcass characteristics
and meat sensory attributes of fattening lambs. Indian J. Anim. Sci. 78, 858–862. Srivastava, J.P., Gupta, B.S., Thakur, S., Verma, A.K., 1990. Utilization of deoiled karanj (Pongamia glabra) cake in kid grower rations. Indian J. Anim. Nutr. 7, 15–20. Vansoest, P.J., Robertson, J.B., Lewis, B.A., 1991. Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J. Dairy Sci. 74, 3583–3597. Vinay, B.J., Kanya, T.C.S., 2008. Effect of detoxification on the functional and nutritional quality of proteins of karanja seed meal. Food Chem. 106, 77–84. Wilson, P.A., Melmed, R.N., Hampe, M.V., Holt, S.J., 1978. Immunochemical study of interaction of soybean trypsin inhibitor with rat intestinal mucosa. Gut 19, 260L 265.
Please cite this article in press as: Rao, S.B.N., Kumar, D.D., Effect of substitution of soybean meal by detoxified karanja cake on diet digestibility, growth, carcass and meat traits of sheep. Small Ruminant Res. (2015), http://dx.doi.org/10.1016/j.smallrumres.2015.03.010