Introduction

Poultry ration consist of cereals and vegetable based protein sources. These ingredients consist of 10-75% of non-starch polysaccharides (NSP) (Choct, 2011). The NSP in cereals form a part of the cell wall structure and in vegetable proteins, especially legumes, play a role as an energy storage material. Poultry being simple stomach cannot digest complex nutrients like non-starch polysaccharides Supplementation of poultry diet with fiber hydrolyzing enzymes does enhance utilization of the complex carbohydrate moiety (Choct, 2006). Because of increase in World’s population and the decline in its food reserve, a more efficient conversion of agro industrial by-products including those rich in NSP, into high quality food is a major area of research today. Soybean meal (SBM) is being used as sole protein source in recent years which contains about 20% NSP (Malathi and Devegowda, 2001). Similarly, other major ingredients used in broiler and layer diets i.e., maize and rice bran contains 9 and 25% NSP, respectively (Malathi and Devagowda, 2001) half of which is cellulose as reported by (Saunders, 1986). The NSPs are insoluble (cellulose) and soluble (β-glucose, arabinoxylan, arabinogalactose, xyloglucon etc). The soluble NSPs have the property to immobilize water in its matrix by forming loose gel network which is responsible for increased viscosity, there by depressing the digestibility of fats, proteins and starch. These NSPs impair activity of endogenous enzymes by reducing the contact intensity between nutrients and enzymes, which results in sticky and moist droppings.

Nowadays, the cost of soybean meal is increasing day by day and because of this the poultry farmers are looking for the alternate protein meals (APM) like- guar meal (GM, protein 33-45%), rapeseed meal (RSM, protein 38-43 %) and cotton seed meal (CSM, protein 40-42%). However, the non-starch polysaccharide (NSP) contents of GM, RSM and CSM are very high (78%, 36.2 and 36.7) respectively. The NSPs can be hydrolyzed with the supplementation of NSP hydrolyzing enzymes. Feed enzymes have been incorporated to improve the nutritive value of poultry diets has become common practice in many countries due to use of feed ingredients containing higher proportion of NSP. However, chicken does not secrete NSP hydrolyzing enzymes to breakdown NSPs present in most of raw materials of plant origin (Close, 1996). Poultry being simple stomached, do not produce enzymes like cellulase, hemicellulase, xylanase and β-glucanase which are required for the digestion of cell wall components of plant material. Inadequate or non-availability of certain enzymes in the digestive tract of chicken reduces their utilization. Enzymes added in the diet break the polymeric chain of fibrous material more effectively, thereby reducing the gut viscosity and improve their nutritive value (Smits and Annison, 1996). Hence to resolve this issue the supplementation of the substrate specific NSP hydrolyzing enzyme combination has been taken as an alternative tool. Hong et al. (2002) found that the use of an enzyme cocktail improved the digestibility of corn-soybean based diets.

Materials and Methods

The present experiment was undertaken on 350 day old ven-cobb straight run broiler chicks consisting of seven treatment groups having ten replicates five birds each. The housing and management of experimental birds was similar to all groups. The experiment was specifically designed keeping in view the replacement of soybean meal as protein source from the broiler diet with the alternate protein sources viz. guar meal (GM), rapeseed meal (RSM) and cotton seed meal (CSM) each at two different levels as alternate protein source was supplemented at 3 (total 9 percent) and 6 percent level (total 18 percent) at Iso-caloric and Iso-nitrogenous levels.

The experiment was a stepping stone to derive a suitable NSP hydrolyzing enzyme combination for the alternate protein sources so as to utilize the low cost alternate protein meals in broiler diet to make the diet economically viable. The pure NSP hydrolyzing enzymes were procured from Advanced Bio- Agrotech Limited, Pune, India. The activity of xylanase, ß-d-glucanase, cellulase, mannase and pectinase was 160000, 200000, 1000000, 200000 IU/g, and 150000 respectively. A total of seven experimental diets were formulated and details are presented in Table 1. The ingredient and nutrient composition of the experimental diets is given in Table 3. However the concentrations of the NSP hydrolyzing enzymes are given in Table 2.The data obtained from the experiment were subjected to appropriate statistical analysis using Statistical Package for Social Sciences (SPSS) software and comparison of means was tested using Duncan’s multiple range tests (Duncan’s, 1955).

Table 1: Details of Broiler Diets

Table 2: Details of the NSP Hydrolyzing Enzyme Concentrations

Table 3: Ingredient Composition of Broiler Diets

¹Trace mineral provided per kg diet: manganese, 120mg; Zinc, 80mg; Iron, 25mg; Copper, 10mg; Iodine,1mg; and Selenium, 0.1mg. ² Vitamin premix provided per kg diet: Vitamin A, 20000IU; Vitamin D₃, 3000IU; Vitamin E, 10mg; Vitamin K,2mg; ³ Riboflavin, 25mg; Vitamin B₁, 1mg; Vitamin B₆, 2mg: vitamin B_12, 40mcg and Niacin, 15mg

Results and Discussion

The body weight gain was significantly higher (P<0.05) in control diet T₁ and 3% APM supplemented with 1X and 2X HC throughout the three phases of experimental period. The 3% APM diet supplemented with NSPHE @ 1X and 2X T₃ and T₄, 6% APM diet supplemented with 1X NSPHE T₆ have shown significantly higher (P<0.05) BWG during the prestarter phase. 3 % APM control diet had shown comparatively better BWG (P<0.05) than that of T₅ and T_7, which have shown significantly poor BWG (Table 4). During the starter phase, the BWG was significantly higher (P<0.05) in T₁, T₃ and T₄ compared with T₂, T_6, T₇ and T₅. The starter phase BWG was significantly lower in the T₅ followed by T_7. The finisher phase BWG was significantly better (P<0.05) amongst the treatment groups T₁, T₂, T₃, T₄ and T₆ compared with T₅ and T₇. The treatment T₅ comprising of 6 % APM control had shown poor BWG during the finisher phase, which was followed by T₇ comprising of 6 % APM + 2X NSPHE (Table 4).

Table 4: Effect of supplementation of non starch polysaccharide hydrolyzing enzymes in diets containing APM (GM,RSM,CSM) each at 3 and 6 % levels on body weight gain(g)of broilers (0-42 days)

Values bearing different superscripts within a columns differ significantly (P<0.05)

The findings are in accordance with Abbas et al. (1998) revealed that NSP enzyme supplementation to fibrous diet improved the growth rate of broilers. Zanella et al. (1999) investigated the effect of a commercial enzyme cocktail containing xylanase, protease and amylase on performance of broilers fed a corn-soybean meal based diet. Enzyme supplementation improved BWG. Hanumantha Rao et al. (2003) reported that the supplementation of enzyme cellulase, xylanase, amylase, protease and phytase individually to maize soybean based diets, significantly (P<0.05) improved BWG of broilers at 42 d. Lazaro et al. (2003) also reported that enzyme supplementation improved performance of birds fed rye lets. Edwin et al. (2004) concluded significant increase in fifth week BWG (P <0.01) of the Japanese quail supplemented with NSP hydrolyzing enzymes. Olukosi et al. (2007) and Gao et al. (2007) reported improved BWG with enzyme supplementation. Cowieson and Ravindran (2008^b) who concluded that enzyme supplementation in broilers fed maize based diets varying in nutrient density improved weight gains of the birds (P<0.001), Berwal et al. (2008) observed feed intake (P<0.05) in broilers fed diets with higher crude fibre levels supplemented with enzyme. Dinani et al. (2010) reported that enzyme supplementation is beneficial for growth performance of growing quails. Mohammad et al. (2010) reported that supplementation of 0.1% (Avizyme) 1100 (xylanase and β-glucanase 300 U/g) to barley based finisher diets improved weight gains. Larhang and Torki (2011) reported that inclusion of different levels of guar meal (0, 4 and 8%) and commercial enzyme (0 and 0.05% of Natuzyme) resulted highest BW and BWG. Narasimha et al. (2013^c) concluded that supplementing sub-optimal energy diets with NSP enzymes along with synbiotics and phytase improved BWG, FCR. Rama Rao et al. (2014) reported that inclusion of GM 200g/kg diet with incremental level of enzyme supplements resulted (P<0.05) increased BWG compared to GM 200g/kg diet without enzyme. Santhi et al. (2014) conducted an experiment to find out the effect of supplementation of exogenous cellulase through feed in turkeys up to eight weeks. Nikam et al. (2016) concluded that supplementing NSP hydrolyzing enzymes to Corn soybean meal based diets has influenced the BWG of the broilers.

However the findings are conflicting compared with Mohammad Akbari Gharaei et al., 2012) who reported that inclusion of GM (0, 3, 6 and 9 %) and β – mannanase enzyme (0 and 0.05 %) were not significant on weight gain. Mishra et al. (2013) reported that live weight of guar korma fed group was inferior to SBM fed ones and that mannanase supplementation did not revert this effect. Narasimha et al. (2013^a) reported that non-starch polysaccharide enzymes with or without prebiotics had no effect on BWG of broilers. The prestarter phase had shown significantly better (P<0.05) feed intake in the group fed with 3 % APM diet supplemented with 1X and 2X NSPHE. However 3% APM containing diet and 6 % APM supplemented with 1X NSPHE had also shown significantly higher (P<0.05) feed intake during prestarter phase and was comparable with T₃, T₄and T₅. During prestarter phase, the feed intake in control diet (T₁) and NSPHE @ 2X (T₇) showed significantly low feed intake. The starter phase showed significant difference (P<0.05) in the feed intake in the 6 % APM control diet (T₅) compared with rest of the diets. The standard control diet (T₁) has also shown significantly higher (P<0.05) feed intake but was statistically comparable with T₂, T₃, T₄, T₇ and T₆ (P<0.05). The diets with 3 % APM (T₂) and 3 % APM with 1X (T₃) and 2 X (T₄) NSPHE and (T₆) had shown comparable feed intake which was significantly higher (P<0.05) compared with (T₇). The diet (T₇) has shown significantly poor feed intake during the starter phase. In finisher phase, the feed intake was significantly higher (P<0.05) among the 3 and 6 % APM fed control (T₂ and T₅) compared with rest of diets i.e. T₁, T₃, T₄, T₆and T₇. The 6 % APM diet supplemented with NSPHE @ 2X (T₇) concentration has also shown significantly higher feed intake (P<0.05) (Table 5).

The results are supported by Nadeem et al. (2005) who reported that by supplementing NSP degrading enzymes at higher concentrations (1X HC and 2X HC) to low energy diets resulted in higher feed intake during overall period compared to basal corn-soy control. Cowieson et al. (2010) observed higher feed intake in low energy deficient corn-soy diet (P<0.01) supplemented with NSPHE alone or in combinations in broilers compared to standard diet. Mirakzehi et al. (2010) reported that diet based on soybean meal which was replaced by enzyme treated RSM (0.03%) had similar overall feed intake to control diet. Ramesh and Chandrasekaran (2011) reported increased feed intake in low nutrient density diets supplemented with exogenous enzymes in Babcock male broiler chicks. Nikam and Reddy (2015) concluded that supplementing NSP hydrolyzing enzymes to Corn soybean meal based diets @ 1X higher concentration has influenced the feed intake of the broilers.

Contrary to the present findings several other workers Narasimha Rao (1998) reported no significant effect of enzyme supplementation. Edwin et al. (2004), Hanna et al. (2008) and Cowieson and Ravindran (2008^a) also reported no effect of exogenous enzyme supplementation on feed intake in broilers.

Table 5: Effect of supplementation of non starch polysaccharide hydrolyzing enzymes diets containing APM (GM,RSM,CSM) each at 3 and 6 % level on phase wise feed intake (g) of broilers (0-42 days)

Values bearing different superscripts within a column differ significantly (P<0.05)

The FCR during the pre starter period was significantly (P<0.05) in birds fed with standard control diet (T₁) (Table 6).The phase wise FCR was comparable among the diets T₃, T₄ and T₆and was found to be significantly better (P<0.05) compared with T₂, T₇ and T₅ .The prestarter phase had shown poor FCR in the diets with 3 and 6 % APM control. However it was statistically comparable with T_7.The starter phase FCR was significantly better (P<0.05) in the treatment T₃compared with other treatments. However it was statistically comparable with T₁, T₂, T₄ and T₅except T₃ the FCR was statistically comparable among all the treatments. The diets T₆ and T₇ had significantly poor FCR during starter phase. The finisher phase FCR was significantly better (P<0.05) in the 3 % APM diet supplemented with NSPHE @ 2X concentration (T₄) compared with rest of the treatments. The diets T₁, T₂, T₃ and T₇ has also shown significant improvement (P<0.05) in FCR compared with T₅ and T_6. The diet T₅ had witnessed poor FCR during finisher phase supplementation of NSP hydrolyzing enzymes at 1X and 2X concentrations has influenced the FCR (P<0.05) during overall period (0-42) days (Table 6). Supplementation of NSP enzymes at 1X and 2X levels influenced the FCR (P < 0.05) during overall period (0-42) days. The findings of this experiment were supported by Saleh et al. (2005) who reported the feed conversion ratio was significantly improved in enzyme fed diets. Meng and Slominski (2005), Wang et al. (2005) also reported similar findings with feed conversion ratio (FCR) of chickens in energy deficient diet. Song et al., 2010 and Mohammad et al., 2010 reported better FCR with enzyme supplementation. Narasimha et al. (2013^a) concluded that FCR improved with addition of NSP enzymes alone or in combination with prebiotics. Rama Rao et al. (2014) reported that inclusion of GM 200g/kg diet with incremental level of enzyme supplements resulted (P<0.05) lower FCR. Santhi et al. (2014) conducted an experiment to find out the effect of supplementation of exogenous cellulase through feed in turkeys up to eight weeks. The findings of the study indicated significantly improved feed efficiency in the enzyme supplemented groups. Nikam et al. (2016) concluded that supplementing NSP hydrolyzing enzymes to Corn soybean meal based diets has influenced the FCR of the broilers reared on sub-optimal diets.

These findings are not in accordance with Naqui and Nadeem (2004), Olukosi et.al. (2007), Garipoglu et al. (2006), Gao et al. (2007) and Berwal et al. (2008)

Table 6: Effect of supplementation of non starch polysaccharide hydrolyzing enzymes diets containing APM (GM,RSM,CSM) each at 3 and 6 % level on phase wise feed efficiency of broilers (0-42 days)

Values bearing different superscripts within a column differ significantly (P<0.05)

Conclusion

The NSP hydrolyzing enzyme combinations consisting of xylanase, ß-D-glucanase, cellulase, mannanase and pectinase @ 1X i.e. 3200, 4000, 1520, 8000, and 3200 IU/kg respectively and @ 2X concentration i.e. 6400, 8000, 3040, 16000 and 6400 IU/kg respectively has resulted in improved performance of the broilers with respect to body weight gain, feed intake and feed conversion ratio when alternate protein meals viz. guar meal (GM),rapeseed meal (RSM) and cotton seed meal (CSM) were incorporated each at 3 and 6 % levels in the broiler diets.

References

1. Abbas W, Khan S H and Sarwar M 1998. Sunflower oil meal as a substitute for soybean meal in broiler rations with or without multi enzyme (kemzyme). Pakistan Veterinary Journal 18(3): 124-129.
2. Berwal R S, Lohan O P and Shiag Zile S 2008 Effect of dietary crude fibre levels and enzyme supplementation on performance and carcass characteristics of broiler chicks Indian Journal of Poultry Science 43:(2)175-178.
3. Choct M 2006 Enzymes for the feed industry: past, present and future World’s Poultry Science 62: 5-16.
4. Choct M 2011 Feed Polysaccharides: Nutritional Roles and Effect of Enzymes presentation given at the IV CLANA CBNA/AMENA, Sao Pedro, SP, Brazil in November 2010 Engormix.com.
5. Close W H 1996 Effect of β-mannanase and protease in corn-soya based diets on broiler performance. Pigs Missets. 12:21-4.
6. Cowieson A J, Bedford M R and Ravindran. V 2010 Interactions between xylanase and glucanase in maize-soy-based diets for broilers. British Poultry Science 51 (2) 246-257.
7. Cowieson A J and Ravindran V 2008^a Effect of exogenous enzymes on maize – based diets varying in nutrient density for broilers: growth performance and digestibility of energy, minerals and amino acids. British Poultry Science 49: 340-346.
8. Cowieson A J and Ravindran V 2008^b Sensitivity of broiler starters to three doses of an enzyme cocktail in maize-based diets. British Poultry Science Vol. 49 (1) 37-44.
9. Dinani O P, Tyagi Promod, Shrivastav A K, Tyagi Praveen 2010 Effect of feeding fermented guar meal vis-à-vis toasted guar meal with or without enzyme supplementation on performance of broiler quail. Indian Journal of Poultry Science 45 (2): 150-156.
10. Duncans 1955Multiple range and multiple F tests.Biometrics 11: 1-42.
11. Edwin S C, Viswanathan K, Mohan B and Purushothaman M R 2004 Effect of supplementation of NSP hydrolyzing enzymes on growth performance of Japanese quails. Indian Journal of Poultry Science 39 (3):241-245.
12. Gao F, Jiang Y, Zhoua G H and Han Z K 2007 The effects of xylanase supplementation on performance, characteristics of the gastrointestinal tract, blood parameters and gut microflora in broilers fed on wheat-based diets. Animal Feed Science and Technology. Pp. 1-12.
13. Garipoglu A V, Saricicek B Z and Kilic U 2006 Effects of the commercial enzyme supplementation to the rations on broiler performance. Asian Journal of Animal and Veterinary Advances 1 (1): 42-48.
14. Hanna A Zakaria, Mohammad H, Jalal A R and Amar S Jabrin 2008 Effect of exogenous enzymes on the growing performance of broiler chickens fed regular corn-soybean based diets and the economics of enzyme supplementation Pakistan Journal of Nutrition 7(4): 534-539.
15. Hanumantha Rao M, Ravinder Reddy V and Ramasubba Reddy V 2003 Effect of commercial enzymes on the performance of broiler. Indian Journal of Poultry Science 38 (3), 291-293.
16. Hong D, Burrows H and Adeola O 2002 Addition of enzymes to starter and grower diets for ducks. Poultry Science 81:1842-1849.
17. Larhang R A and Torki M 2011 Evaluating Performance of Broilers Fed Guar Meal- included diet Supplemented by Enzyme. Researches of the first International conference (Babylon and Razi universities) Animal Science Department, Agriculture faculty,Razi University, Kermanshah, Iran.
18. Lazaro R, Gracia M, Edel P and Mateas G G 2003 Influence of enzymes on performance and digestive parameters of broilers fed rye – based diets. Poultry Science 82: 132-40.
19. Malathi V and Devegowda G 2001 In vitro evaluation of non starch polyaccharide digestibility of feed ingredients by enzymes. Poultry Science 80(3): 302-305.
20. Meng X and Slominski B A 2005 Nutritive values of corn, soybean meal, canola meal, and peas for broiler chickens as affected by a multicarbohydrase preparation of cell wall degrading enzymes. Poultry Science 84: 1242-1251.
21. Mirakzehi M T, Tahmasbi A and Ghazi A 2010 The Effect of Different Treatments of Rapeseed Meal on Nitrogen Digestibility and Metabolizable Energy in Broilers and Chicks Performance Journal of Animal and Veterinary Advances 9(19):2517-2521.
22. Mishra A, Sarkar S K, Ray S and Haldar S 2013 Effects of partial replacement of soybeanmeal with roasted guar korma and supplementation of mannanase on performance and carcass traits of commercial broiler chickens. Veterinary World 6(9): 693-697.
23. Mohammad T Farran, George W Barbour, Nada N Usayron, Ali H Darwish, hasan H amatchhlals, Milan Hruby and Vahe M Askarian 2010 Performance and carcass quality of broiler chickens fed a corn-soybean meal diet containing graded barley levels without or with enzymes. Journal of Poultry Science 47: 34-40.
24. Mohammad Akbari Gharaei, Behrouz Dastar, Alireza Hesabi Nameghi, Gholamreza Hashem Tabar and Mahmoud Shams Shargh 2012 Effects of Guar meal with and without β- mannanase enzyme on performance and immune response of broiler chicks. Internatioanl Res. Journal of Applied and Basic Sciences 3 (13) 2785-2793.
25. Nadeem M A, Anjum M I Khan A G and Azim A 2005 Effect of dietary supplementation of non-starch polysaccharide degrading enzymes on growth performance of broiler chicks. PakistanVeterinary Journal 25 (4): 2005.
26. Naqui L U and Nadeem A 2004 Bio availability of metabolisable energy through enzyme supplementation in broiler rations. Pakistan Veterinary Journal 24(2):183-188.
27. Narasimha J, Nagalakshmi D, Ramana Reddy Y and Viroji Rao S. T 2013^a
    Synergistic effect of non starch polysaccharide enzymes, synbiotics and Phytase on performance, nutrient utilization and gut health in broilers fed with sub-optimal energy diets Veterinary World, 6(10): 754-7601.
28. Narasimha, J. Nagalakshmi D., Ramana Reddy Y. and. Viroji Rao S.T 2013^c Screening of non starch polysaccharide degrading enzymes and enzyme combinations for their ability to degrade non starch polysaccharides of broiler diet by two stage in vitro digestion assay Indian Journal of Poultry Science (2013) 48(1): 1-5.
29. Narsimha Rao 1998 Effect of enzymes in feed on broiler performance. M V Sc. Thesis submitted to ANGRAU Rajendranagar, Hyderabad.
30. Nikam M. G. and Ravinder Reddy V. 2015 Effect of supplementation of Non Starch Polysaccharide Hydrolyzing Enzymes on Performance of Broilers International Journal of Science and Research (IJSR) : 4 (12):1690-1693.
31. Nikam M. G. Ravinder Reddy V,Raju M.V.L.N., Kondal Reddy K and Narasimha J 2016 Effect of dietary supplementation of non starch polysaccharide hydrolyzing enzymes in broilers reared on corn-soya meal standard and basal diet. International Journal of Agricultural Science and Research (IJASR) 6 (3):389-396.
32. Olukosi O A, Cowieson A J and Adeola O 2007 Age-related influence of a cocktail of xylanase, Amylase and Protease or Phytase individually or in combination in broilers. Poultry Science 86: 77-86.
33. Rama Rao S V, Prakash B, Raju M V L N, Panda A K and Murthy O K 2014 Effect of supplementing non-starch polysaccharide hydrolyzing enzymes in guar meal based diets on performance, carcass variables and bone mineralization in vanaraja chicken. Animal Feed Science Technology 188: 85-91.
34. Ramesh J and Chandrasekaran 2011 effect of exogenous enzyme supplementation on performance of cockerels, Tamil Nadu Journal of Veterinary and Animal Sciences 7(1): 29-34.
35. Saleh F, M Yamamoto, M Tahir, A Ohtsuka and K Hayash 2005 A new natural feed additive for broiler chickens. Poult. Sci. Asso. Annual Meeting. Edmonton, Canada, pp: 36-54.
36. Santhi, D.,Thyagarajan,D and Ramesh,J. 2014. Effect of exogenous cellulase supplementation in feed on turkey poult performance. Indian Veterinary Journal 91(02) 9-11.
37. Smits C H M and Annison G 1996 Non starch plant polysaccharides in broiler nutrition towards a physiologically valid approach to their determination. World Poultry Science Journal 52: 203-221.
38. Song Xizo Zhen, Zeng Fuhai, yang Xiujiang, Qu Ming Ren and Zhang Wei 2010 Effects of different combinations of NSP enzyme preparations on small intestine mucosal structure and nutrient apparent availability of 1-21 days old broilers. Chinese Journal of Animal Nutrition 22 (6): 1730-1737.
39. Wang Z R, Oiao S Y, Lu W O and Li D F 2005 Effects of enzyme supplementation on performance, nutrient digestibility, gastrointestinal morphology and volatile fatty acid profiles in the hindgut of broilers fed wheat-based diets. Poultry Science 84: 875-881.
40. Zanella I, Sakomura N K, Silversides F G, Fiqueirdo A and Oack M 1999 Effect of enzyme supplementation of broiler diets based on corn and soybeans. Poultry Science ,78: 561- 568.