The Indian Journal of Small Ruminants 2013, 19(1): 45-49

Indian Journal of Small Ruminants

NUTRIENT UTILIZATION IN SHEEP FED SORGHUM STOVER-BASED DIETS SUPPLEMENTED WITH CELLULASE AND XYLANASE T. Vijay Bhasker, D. Nagalakshmi*, D. Srinivasa Rao and T. Raghunandan Department of Animal Nutrition, College of Veterinary Science, Rajendranagar, Hyderabad- 500 030, Andhra Pradesh *E-mail address: dnlakshmi@rediffmail.com
Manuscript received on 11.06.2012, accepted on 09.08.2012

ABSTRACT
An experiment was conducted on 12 male lambs (6-8 months of age, 20.141.76 kg body weight) fed sorghum stover as sole roughage source in total mixed ration (TMR) (roughage: concentrate, 50:50) to study the effect of supplementing exogenous fibrolytic enzymes (EFE) comprising of 38400 cellulase and 25600 xylanase IU /g DM(dry matter) stover on nutrient utilization. The required amount of EFE was added to concentrate mixture of TMR diet before blending with chopped stover. The animals were randomly allotted to 2 dietary groups i.e., sorghum stover based TMR and the TMR supplemented with EFE. They were fed with respective diets for a preliminary period of 21 days followed by 7 days collection period. Supplementation of EFE improved the digestibility of organic matter, nitrogen free extract, cell contents, neutral detergent fiber and cellulose in sheep, with no effect on crude fiber and ether extract digestibilities. Though non-significant, crude protein digestibility lowered with EFE supplementation resulting in lower digestible crude protein content of diet. The total digestible nutrient and metabolic energy content improved with EFE supplementation. No effect of EFE supplementation was observed on calcium and phosphorus balances. The lower nitrogen intake with EFE supplementation resulted in lower nitrogen balance, but when expressed as per cent intake, the nitrogen balance did not vary. The results of this study indicated the fibrolytic enzymes supplemented to sorghum stover-based rations at concentration of 38400 IU cellulase and 25600 xylanase IU/g DM stover improved the nutrient digestibility and TDN content. Key words: Exogenous fibrolytic enzymes, Nutrient utilization, Sheep, Sorghum stover

n tropical countries like India, small ruminants are traditionally managed on grazing, but in view of continuous depletion of grazing lands and scarcity of feeds and fodder, farmers are forced to utilize the crop residues like straws, stovers and other agro-industrial byproducts for feeding, which are high in fibre. To increase the nutrient availability from these crop residues and poor quality roughages, nowadays addition or supplementation of exogenous fibrolytic enzymes as feed additives has received considerable attention. Exogenous fibrolytic enzymes work in synergy with the endogenous rumen microbiological enzymes to enhance the digestibility and nutritive value of high fibrous diet (Morgavi et al., 2000). The in vitro dry matter (DM) and neutral detergent fiber (NDF) degradability from hays, hay-based total mixed rations

(TMR) and silages improved with exogenous fibrolytic enzyme (EFE) supplementation (Gallardo et al., 2010). Many researchers have reported improvement in nutrient digestion with addition of EFE to concentrate portion of TMR due to adequate preingestion enzyme substrate complex interaction (Krueger and Adesogan, 2007). Most of the research carried in developed countries on EFE supplementation was on low fiber roughages or high concentrate diets. The action of enzymes is target specific (Pinos Rodriguez et al., 2002) and hence the enzyme combination developed for concentrate or silage-based diets might not be effective for straw based diets. Thus, the present experiment was aimed to test fibrolytic enzyme cocktail in sheep in terms of nutrient utilization and balances.

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MATERIALS AND METHODS The cellulase and xylanase enzymes used in the study had activities of 1x106 and 1.6x106 IU/g, respectively and were active in alkaline pH. A total mixed ration was formulated with sorghum stover as sole roughage source in roughage concentrate ratio of 50:50 (TMR-SS). The sorghum stover was chopped and mixed with concentrate mixture which consisted of 35% groundnut cake, 40% maize, 22% de-oiled rice bran, 2% salt and 1% mineral mixture and vitamin AD3 (20g/qt). The experimental ration was the TMR-SS to which fibrolytic enzyme cocktail (cellulase+xylanase, 38400-25600 IU/g sorghum stover) was supplemented (TMR-SS +EFE). The calculated quantity of enzyme was accurately weighed and mixed in the required concentrate mixture and then mixed with chopped sorghum stover manually for about 10 minutes before feeding to sheep daily. Twelve Deccani male weaners (6 to 8 months) with an average body weight of 20.141.76 kg were randomly distributed into 2 groups, one group was fed with control TMR (TMR-SS) and the other group on TMR supplemented with EFE (TMR-SS + EFE). The respective rations were offered twice daily at 9.00 am and 3.00 pm to meet the nutrient requirements (ICAR, 1998). The dry matter intakes of un-supplemented and enzyme-supplemented sorghum stover basedTMR were 560.2 g/day and 338.2 g/day respectively. The leftover was weighed on the next day morning before cleaning and feeding. Clean, fresh and wholesome

water was made available to each group of animals at all times. At the end of preliminary feeding period of 21 days, a digestion-cum-metabolic trial of 7 days inclusive of 2 days acclimatization period was conducted to assess the digestibility of nutrients and balances of nitrogen, calcium and phosphorus. Animals harnessed with faecal collection bags were kept in well-ventilated individual metabolic cages with feeding and watering facilities during the trial. Daily record of feed offered, left over, faeces and urine voided for 24 h was recorded. Representative samples of feed offered, leftover, faeces and urine voided were collected daily to arrive at the nutrient digestibility and balances. Feed ingredients, feed offered, feed refusal, faecal samples and urine samples were analyzed for proximate constituents (AOAC, 1997), fibre fractions (Van Soest et al., 1991), calcium (Talpatra et al., 1940) and phosphorus (AOAC, 1997).The data were subjected to one-way ANOVA and comparison of means was tested using students 't' test (Snedecor and Cochran, 1989). RESULTS AND DISCUSSION The sorghum stover used in the present study contained 3.62% crude protein (CP) and 50.03% crude fire (CF) and was comparable to that reported by (Hamed and Eliman, 2009). The TMR-SS contained 10.49% CP, 47.11% acid detergent fibre (ADF) and 35.76% cellulose. Supplementation of EFE did not affect the chemical composition of TMR (Table 1).

Table 1. Chemical composition (on %DM basis) of experimental diets, sorghum stover and concentrate mixture Nutrient Organic matter Crude protein Crude fibre Ether extract Nitrogen free extract Total ash Neutral detergent fibre Acid detergent fibre Hemicellulose Cellulose Lignin Calcium Phosphorus 46 Total mixed ration (TMR) TMR - SS TMR - SS + EFE 87.52 88.75 10.49 10.75 30.53 29.29 2.47 2.51 44.02 46.20 9.48 8.39 74.50 74.10 47.11 48.91 27.39 25.18 35.76 39.12 9.70 8.69 0.78 0.80 0.83 0.80 Sorghum stover 85.10 3.62 50.03 1.01 30.46 9.59 78.47 49.67 28.80 37.35 11.67 0.76 0.53 Concentrate mixture 85.36 15.78 8.46 3.41 57.8 7.47 40.28 21.12 19.16 14.17 5.47 1.89 1.95

The Indian Journal of Small Ruminants 2013, 19(1): 45-49

Fibrolytic enzymes on nutrient utilization of sorghum stover in sheep

Supplementing EFE to sorghum stover-based TMR had no significant affect on DM digestibility, while organic matter (OM) (P<0.05) and nitrogen free extract (NFE) (P<0.01) digestibilities significantly increased with EFE supplementation (Table 2). Though, non-significant, the EE and CF digestibilities improved by 7.35 and 8.32%, respectively. Similarly, Titi and Tabba (2004) reported higher OM and CF digestibility in lambs, calves and replacement heifers with supplementation of cellulase (200 units /mg) at 1.50 g/tonne of wheat straw in TMR. An enhancement in CF digestibility was observed in Murrah buffaloes with fibrolytic enzymes (cellulase 4000 IU and xylanase 7900 IU/g) supplemented at 1.5 g to wheat

straw-based TMR (Chandra Shekhar et al., 2010). Bala et al. (2009) reported improved digestibilities of DM and OM in goats fed wheat straw-based diets fortified with cellulase and xylanase @18000 and 18,750 IU/kg. While no effect of supplementing EFE was observed in lambs supplemented with 3 g (Avellaneda et al., 2009) to guinea grass and 4 g (Reddish and Kung, 2007) to commercial diet, which might be due to lower concentration of enzymes. Though, non-significant CP digestibility lowered by 10.87% in enzyme-supplemented group, might be due to its lower intake. On the contrary, CP digestibility was not affected with EFE supplementation (Titi and Tabba, 2004; Avellaneda et al., 2009).

Table 2. Digestibility of proximate constituents (%) in ram lambs fed sorghum stover based TMR supplemented with EFE Attribute Proximate constituent Dry matter Organic matter Crude protein Ether extract Crude fibre Nitrogen free extract Fibre fraction Cell contents Neutral detergent fibre Acid detergent fibre Hemicellulose Cellulose 70.662.68A 45.602.22b 40.732.25 67.711.62 54.854.08
B

TMR-SS 51.822.00 60.812.50

b

TMR -SS+EFE 53.951.75 67.821.45

a

SEM 1.307 1.738 2.060 2.060 2.558 1.751 2.771 1.889 1.732 1.173 2.858

P value 0.443 0.036 0.071 0.071 0.105 0.003 0.010 0.014 0.059 0.384 0.010

62.773.08 82.053.41 37.294.31 70.151.78

A

55.953.62 89.411.29 45.611.82 79.261.42

A

57.653.11B 54.171.85a 47.142.01 65.561.72 68.321.09

A

Means bearing different superscripts in a row differ significantly (P<0.01; P<0.05), SS: Sorghum stover, EFE: Exogenous Fibrolytic Enzyme, TMR: Total Mixed Ration

Significant increase (P<0.05) in NDF and cellulose digestibility was observed in sheep with EFE supplementation, while the ADF digestibility nonsignificantly improved by 6.41% (Table 3). Such positive effect on fiber digestibility with EFE addition was reported in buffalo calves (Chandra Shekhar et al., 2010) and sheep (Titi and Tabba, 2004) fed wheat straw-based TMR, finishing steers fed alfalfa hay and corn stover based TMR (Carreon et al., 2010) and sheep fed grass hay-based diet (Giraldo et al., 2008).

The increase in the OM and fibre digestibilities observed in the present study might be due to the enhanced attachment of rumen microorganisms to the feed particles and synergistic effect of the exogenous enzymes with the hydrolases of the ruminal microorganisms (Morgavi et al., 2000). Also addition of EFE to concentrate portion of TMR, might have given scope for adequate pre-ingestion enzyme substrate complex interaction, resulting in higher nutrient digestibility (Krueger and Adesogan, 2007).

The Indian Journal of Small Ruminants 2013, 19(1): 45-49

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Table 3. Nutritive value of sorghum stover sorghum stover based TMR supplemented with EFE Diet DCP (%) TDN (%) DE (M cal/kg) ME (M cal/kg) TMR-SS 6.850.36
a B

TMR -SS+EFE 5.530.35

b A

SEM 0.313 2.939 0.106 0.106

P value 0.026 0.001 0.001 0.001

54.102.39 2.390.11

71.141.80 3.140.08 2.570.30

1.960.04

Means bearing different superscripts in a sub- column differ significantly (P<0.01; P<0.05)

The improvement in OM, NFE, EE and fiber digestibilities with EFE supplementation resulted in higher energy content (TDN, DE, ME) of diet. El-Kady et al. (2006) reported significant increase in nutritive value of sugar beet pulp-based ration by adding EFE (cellulase, xylanase, -amylase and polyglacturenase activities @ 114.2, 662.1, 1065.1 and 216.2 IU/g fermented SBP). Though non- significant, the CP digestibility lowered by 10.87 % resulting in lower (P<0.05) DCP content of diet. All the lambs were on positive nitrogen, calcium and phosphorus balance indicating that both the diets supplied required quantity of respective nutrients and the nutrients absorbed were fully utilized by the animal (Table 4). There was no significant difference in nutrient balance and nutrient retention as percent of intake in case of calcium and phosphorus upon EFE supplementation

(Table 4). The nitrogen intake was lower (P<0.01) in experimental group which was due to lower (P>0.05) feed intake with EFE supplementation and the lower nitrogen intake resulted in lower nitrogen balance compared to un-supplemented lambs. But nitrogen balance when expressed as percent intake was comparable between EFE supplemented and un-supplemented lambs. These findings were in accordance with the finding of Avellaneda et al. (2009), who observed no effect of EFE supplementation (100 xylose units/ g; 3g/ day/ lamb) on nitrogen balance in lambs fed on guinea grass hay. Similarly, Knowlton et al. (2002) reported that no significant difference in nitrogen balance in early and late lactating cows fed TMR containing 45% and 61% forage, respectively supplemented with 204 g enzyme/ tone DM (15000 units/g cellulase) compared to un-supplemented ones.

Table 4. Nutrient balance (g) in lambs fed sorghum stover based TMR supplemented with EFE Diet Nitrogen Intake Outgo Faeces Urine Balance Retention as % of intake Calcium Intake Outgo Faeces Urine Balance Retention as % of intake Phosphorus Intake Outgo Faeces Urine Balance Retention as % of intake TMR-SS 9.780.336A 3.592.34A 3.350.52a 2.630.37a 26.813.60 4.240.163 2.280.09A 0.150.01B 1.81 0.22 37.773.97 5.110.23 2.190.24A 0.340.07 2.150.35 41.326.18 TMR -SS+EFE 5.360.593B 2.340.28B 1.640.41b 1.390.20b 25.792.85 2.51 0.277 1.080.19B 0.160.01A 1.260.26 26.673.95 4.640.49 0.850.18B 0.410.06 1.620.30 34.353.97 SEM 0.740 0.249 0.422 0.274 2.194 0.254 0.284 0.207 0.171 3.150 0.267 0.247 0.046 0.234 3.655 P value 0.001 0.004 0.016 0.014 0.829 0.798 0.001 0.001 0.114 0.075 0.406 0.001 0.449 0.281 0.365

Means bearing different superscripts in a row differ significantly (P<0.01; P<0.05) 48 The Indian Journal of Small Ruminants 2013, 19(1): 45-49

Fibrolytic enzymes on nutrient utilization of sorghum stover in sheep

From the present study, it can be concluded that supplementation of EFE (38400 cellulase- 25600 xylanase IU/g DM of sorghum stover) to sorghum stove- based TMR improved the organic matter and fiber digestibility in sheep with no affect on Ca and P balances. It is only a 21 days experiment, followed by metabolism trial and needs further long-term study to see the role of enzyme supplementation on production performance. REFERENCES
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Giraldo, L.A., Tejido, M.L., Ranilla, M.J., Ramos, S. and Carro, M.D. 2008 Influence of direct-fed fibrolytic enzymes on diet digestibility and ruminal activity in sheep fed a grass hay-based diet. Journal of Animal Sciences 86: 1617-1623. Hamed, A.H.M. and Eliman, M.E. 2009. Effects of chopping on utilization of sorghum stover by Nubian goats. Pakistan Journal of Nutrition 8: 1567-1569. ICAR. 1998. Nutrient Requirements of Sheep. Indian Council of Agricultural Research, New Delhi, India. Krueger, N.A. and Adesogan, A.T. 2007. Effect of different mixtures of fibrolytic enzymes on digestion and fermentation of bahia grass hay. Animal Feed Science and Technology 145: 84-94. Morgavi, D.P., Beauchemin, K.A., Nsereko, V.L. and Rode, L.M. 2000. Synergy between ruminal fibrolytic enzymes and enzymes from Trichoderma longibrachiatum in degrading fiber substrates. Journal of Dairy Science 83: 1310-1321. Pinos-Rodrguez, J.M., Gonzalez, S.S., Mendoza, G.D., Barcena, R., Cobos, M.A., Hernandez, A. and Ortega, M.E. 2002. Effect of exogenous fibrolytic enzyme on ruminal fermentation and digestibility of alfalfa and rye-grass hay fed to lambs. Journal of Animal Science 80: 3016-3020. Reddish, M.A. and Kung, L. Jr. 2007. The effect of feeding a dry enzyme mixture with fibrolytic activity on the performance of lactating cows and digestibility of a diet for sheep. Journal of Dairy Science 90: 4724-4729. Snedecor, G.W. and Cochran, W.G. 1989. Statistical Methods 8th edn., Iowa State University press, Ames, Iowa, USA. Talapatra, S.K., Roy, S.C. and Sen, K.C. 1940. The analysis of mineral constituents in biological materials. Estimation of phosphorus, chlorine, calcium, manganese, sodium and potassium in foodstuffs. Indian Journal of Veterinary Science and Animal Husbandry 10: 243. Titi, H.H. and Tabba, M.J. 2004. Efficacy of exogenous cellulase on digestibility in lambs and growth of dairy calves. Livestock Production and Science 87: 207-214. Van Soest, P.J., Robertson, J.B. and Lewis, B.A. 1991. Methods for dietary fiber, neutral detergent fiber and non- starch polysaccharides in relation to animal nutrition. Journal of Dairy Science 74: 3583-3597.

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