The Effect of Protease Enzyme and Replacement Soybean Meal by Cotton Seed Meal on Protein Efficiency and Performance in Broiler Chickens

Iranian Journal of Applied Animal Science

دوره 13، شماره 1، خرداد 2023، صفحه 143-151   

نوع مقاله: Research Articles

نویسندگان

M. Safari؛ A.A. Saki^* ؛ S. Mirzaie Goudarzi؛ A. Ahmadi؛ A. Ashoori

Department of Animal Science, Faculty of Agriculture, Bu Ali Sina University, Hamedan, Iran

چکیده

This study was conducted to investigate the effect of replacing soy bean meal (SBM) by cottonseed meal (CSM) with or without of protease enzymes, on performance, in-vitro protein digestibility, carcass characteristics, gastrointestinal morphology and hematological parameters in broilers. A total 450 one-day-old mixed sex broiler chickens Ross 308 were managed in 6 treatments, 5 replicates and 15 chickens in each in a 2 × 3 factorial arrangement with completely randomized design (CRD). Treatments consist of: treatment 1 (0% CSM, -protease), treatment 2 (0% CSM, +protease), treatment 3 (20% CSM, -protease), treatment 4 (20% CSM, +protease), treatment 5 (30% CSM, -protease), treatment 6 (30% CSM, +protease). The results have shown that there were no significant affected on feed intake, body weight gain and feed conversion ratio by various treatments in starter period (P>0.05). Digestive organs and body weight were not affected by treatments at 27 days of age. Breasts weight was significantly increased by 20% replacement SBM with CSM (P<0.05). Protein digestibility was increased significantly by protease enzymes and in vitro method. Serum cholesterol levels reduced by enhanced levels of replacement CSM (P<0.05). There was concluded that no adverse effects were found by replacing 30% SBM with CSM on performance status, as well as improved in vitro protein digestibility.

کلیدواژه‌ها

broiler chicken؛ cottonseed meal؛ protease enzyme؛ soy bean meal

اصل مقاله

INTRODUCTION

Soybean meal (SBM) is a predominant protein source in formulation poultry diets. When the soybean meal price increases, poultry producers seek some unconventional protein sources such as cottonseed meal used as a replacement by soybean meal in poultry diet (Liu et al. 2006). Now in regarding to soybean meal price in poultry industry most of researchers in throughout the world try to identified and find out an alternative protein source. However, it is difficult to have a consistent supply of good quality soybean meal in poultry production at a range of reasonable price (Xiong et al. 2016). Cottonseed meal (CSM) is a by-product of process to extract oil from cotton seeds arid contains 40 or 44% crude protein depending on the extent of hull separation prior to the oil extraction process (Heidarinia and Malakian, 2011). It is a potential alternative of SBM in poultry diet (Nagalakshmi et al. 2007). When broilers were fed cottonseed meal based diets, the presence of anti-nutritional factors such as gossypol disturbed the absorption of nutrients and reduces amino acid digestibility. This is may increase the need for protease enzyme demand to more nutrient digestibility and optimize the growth in poultry (Blevins et al. 2010). Also, high demands of amino acids by the broiler chicken could be met by increasing protein contents or added exogenous protease enzyme (Nazemzadeh et al. 2017). On the other hand, Gossypol, a potentially toxic agent, has limited the use of cottonseed meal, as an attractive protein source for poultry feed. The toxic effects of gossypol may be alleviated by supplementation of soluble iron salts that could be bind by gossypol (Shawrang et al. 2011). Enzyme supplementation has become increasingly popular in animal feeds in recent years, the most commonly used are proteases (Kaczmarek et al. 2014). Consequently, it has been suggested that the immature of the digestive system in young chickens may result in the poor utilization of dietary nutrients (Jin et al. 1998). The nutrient digestion rather than the ability of absorption has been indicated to be the primary limiting factor (Parsons, 2004). Based on SBM price, this study was focus to CSM as an alternative protein source and protease enzyme on nutrient digestibility and broiler performance. This study aimed was to investigate the effect of replacing soy bean meal (SBM) by cottonseed meal (CSM) with or without of protease enzymes, on performance, in vitro protein digestibility, carcass characteristics, gastrointestinal morphology and hematological parameters in broilers.

 

MATERIALS AND METHODS

The experiment was conducted at poultry farm in the Faculty of Agricultural in, Bu-Ali Sina University, Hamedan, Iran. A total 450 one-day-old mixed sex broiler chickens Ross 308 with initial body weight of 38.86 ± 7.2 gram were designed by 6 treatments, 5 replicates and 15 chickens in each in a 2 × 3 factorial arrangement with completely randomized design (CRD). Treatments includes: treatment 1 (0% CSM, without protease), treatment 2 (0% CSM, with protease), treatment 3 (20% CSM, without protease), treatment 4 (20% CSM, with protease), treatment 5 (30% CSM, without protease), treatment 6 (30% CSM, with protease). Ferrous sulfate (0 or 200 g/ton) was used for disable gossypol cottonseed meal and protease enzyme 200 g/ton (ton; 75000 PROT/g; Ronozyme® ProAct) in the diet. Experimental diets were considered in starter, grower and finisher periods (1 to 10, 11 to 24 and 25 to 42 days of age respectively). Diets were formulated according to Ross 308 recommend in 2014. All of the diets were formulated based on standardized ileal digestible amino acids (SID) and ideal protein (Table 2, 3 and 4). Diet, in mash form, and water were provided in ad libitum consumption. The feed intake, body weight and feed conversion ration were considered on a weekly period.

 

Chemical analysis

The dry matter, crude (ash, fiber, protein and fat) content of corn, corn gluten meal (CGM), SBM and CSM were analyzed in the Feed Analysis Laboratory using proposed by the Association of Official Analytical Chemists, AOAC methods (AOAC, 2000) as shown in Table 1. Acid detergent fiber (ADF), Neutral detergent fiber (NDF) and the amino acid profile of corn, CGM, SBM and CSM were analyzed respectively by method of Van Soest et al. (1991) and Near Infrared Reflectance Spectroscopy (NIRs) (Table 1). The exogenous protease enzyme was used (Ronozyme ® ProAct) produced by nutritional products DSM, Parsippany, NJ, USA and the commercial product had a protease activity of 75.000 U/g and the unit of activity was defined as the amount of enzyme that released 1 µmol of p-nitroaniline from 1 mM substrate (Suc-Ala-Ala-Pro-Phe-pNA) per minute at pH 0.9 and temperature 37 ˚C.

 

Statistical analysis

A completely randomized design (CRD) was applied. Data were analyzed by the GLM procedure (SAS, 2004). Means were separated by Duncan’s multiple range tests. Treatments differences were considered significant at (P<0.05).

 

RESULTS AND DISCUSSION

The results of broiler chickens performance were presented in Table 6. There was no significant effect on performance parameters in the entire period by interaction between cottonseed meal and protease enzyme (P>0.05). Improved feed conversion ratio (FCR) was found by protease enzyme in broiler diet. The feed conversion ratio was significantly lower by interaction between CSM and protease enzyme in comparison without enzyme treatment (P=0.053). In addition, the European broiler index (EBI) increased by control diet and 20% replacing SBM with CSM without enzymes (P<0.05). As shown in Tables 6 and 7, no significant differences were found on relative weight of carcass, breast, leg, heart, liver, bursa of Fabricius, between treatments (P>0.05). Proventriculus and pancreas weight were significantly increased by 30% replacing SBM with CSM compared with control and 20% replacing (P<0.05).The highest weight of gizzard was observed by 30% replacing SBM with CSM compared with control diet (P<0.05). In vitro dry matter digestibility was decreased with higher levels of dietary CSM (P<0.05). Protein digestibility were increased by 30% replacing SBM with CSM with enzyme in comparison to all treatment’s groups (P<0.05) (Table 8). As shown in Table 8, no significant differences were observed in the albumin and total protein in blood plasma of broiler chicken in different treatments (P>0.05).

 

Table 1 Chemical compositions of dietary ingredients (%; as-is)

¹ SID amino acid measured by NIRS.

NDF: neutral detergent fiber; ADF: acid detergent fiber; GCM: gluten corn meal; SBM: soy bean meal and CSM: cotton seed meal.

 

Table 2 Components of experimental diets and their chemical analysis (%, as-is basis) (starter period from 1 to 10 days)

¹ 1: Control without enzyme; 2: Control with enzyme; 3: 20% replacing CSM with SBM without enzyme; 4: 20% replacing CSM with SBM with enzyme; 5: 30% replacing CSM without SBM without enzyme and 6: 30% replacing CSM with SBM without enzyme.

² SBM: soybean meal; GCM: gluten corn meal; CSM: cottonseed meal; DCP: dicalcium phosphate and NaCl: common salt.

³ Mineral premixes supplied per kilogram of diet: Mn (MnSO₄·H₂O): 66 mg; Zn (ZnO): 66 mg; Fe (FeSO₄·H₂O): 33 mg; Cu (CuSO₄·5H₂O): 6 mg; I: 0.9 mg; Co (CoSO4·H2O): 0.2 mg and Se (Na₂SeO₃): 0.2 mg.

⁴ Vitamin premixes supplied per kilogram of diet: vitamin A (retinyl acetate): 10000 IU; vitamin D₃: 2750 IU; vitamin E (α-tocopheryl acetate): 20 IU; Menadione: 3.0 mg; Thiamin: 2.5 mg; Riboflavin: 6.0 mg; Pyridoxine: 2.5 mg; vitamin B₁₂: 12 µg; Folic acid: 1.5 mg; Niacin: 20 mg; Ca-pantothenate: 15 mg and Biotin: 80 µg.

⁵ AMEn: apparent metabolizable energy corrected for nitrogen; CP: crud protein; Ca: calcium; AP: available phosphate and DM: dry matter.

 

Table 3 Components of experimental diets and their chemical analysis (%, as-is basis) (from 11 to 24 days)

¹ 1: Control without enzyme; 2: Control with enzyme; 3: 20% replacing CSM with SBM without enzyme; 4: 20% replacing CSM with SBM with enzyme; 5: 30% replacing CSM without SBM without enzyme and 6: 30% replacing CSM with SBM without enzyme.

² SBM: soybean meal; GCM: gluten corn meal; CSM: cottonseed meal; DCP: dicalcium phosphate and NaCl: common salt.

³ Mineral premixes supplied per kilogram of diet: Mn (MnSO₄·H₂O): 66 mg; Zn (ZnO): 66 mg; Fe (FeSO₄·H₂O): 33 mg; Cu (CuSO₄·5H₂O): 6 mg; I: 0.9 mg; Co (CoSO4·H2O): 0.2 mg and Se (Na₂SeO₃): 0.2 mg.

⁴ Vitamin premixes supplied per kilogram of diet: vitamin A (retinyl acetate): 10000 IU; vitamin D₃: 2750 IU; vitamin E (α-tocopheryl acetate): 20 IU; Menadione: 3.0 mg; Thiamin: 2.5 mg; Riboflavin: 6.0 mg; Pyridoxine: 2.5 mg; vitamin B₁₂: 12 µg; Folic acid: 1.5 mg; Niacin: 20 mg; Ca-pantothenate: 15 mg and Biotin: 80 µg.

⁵ AMEn: apparent metabolizable energy corrected for nitrogen; CP: crud protein; Ca: calcium; AP: available phosphate and DM: dry matter.

 

Also, plasma cholesterol levels of broiler were decreased significantly by 30 and 20% SBM replacement with CSM compared with control diet (P<0.05). No significant effect was found on villus height, villus thickness, villus height to crypt depth ratio in the jejunum by different levels replacement of SBM with CSM and protease enzyme in the diet (P>0.05) (Table 9). Results of this study have shown there is no significant differences in performance by treatments. Similarly, Azman and Yilmaz (2005) have reported 20% replacing CSM with SBM no significant differences in body weight, daily weight gains, daily feed intake and feed conversion ratios. Cotton seed meal, in spite of undesirable properties of gossypol, was one of the protein sources that could be used as replacement with soybean meal in diet without any adverse effects on the performance efficiency of broiler chickens (Tang et al. 2012).

 

Table 4 Components of experimental diets and their chemical analysis (%, as-is basis) (from 25 to 42 days)

¹ 1: Control without enzyme; 2: Control with enzyme; 3: 20% replacing CSM with SBM without enzyme; 4: 20% replacing CSM with SBM with enzyme; 5: 30% replacing CSM without SBM without enzyme and 6: 30% replacing CSM with SBM without enzyme.

² SBM: soybean meal; GCM: gluten corn meal; CSM: cottonseed meal; DCP: dicalcium phosphate and NaCl: common salt.

³ Mineral premixes supplied per kilogram of diet: Mn (MnSO₄·H₂O): 66 mg; Zn (ZnO): 66 mg; Fe (FeSO₄·H₂O): 33 mg; Cu (CuSO₄·5H₂O): 6 mg; I: 0.9 mg; Co (CoSO4·H2O): 0.2 mg and Se (Na₂SeO₃): 0.2 mg.

⁴ Vitamin premixes supplied per kilogram of diet: vitamin A (retinyl acetate): 10000 IU; vitamin D₃: 2750 IU; vitamin E (α-tocopheryl acetate): 20 IU; Menadione: 3.0 mg; Thiamin: 2.5 mg; Riboflavin: 6.0 mg; Pyridoxine: 2.5 mg; vitamin B₁₂: 12 µg; Folic acid: 1.5 mg; Niacin: 20 mg; Ca-pantothenate: 15 mg and Biotin: 80 µg.

⁵ AMEn: apparent metabolizable energy corrected for nitrogen; CP: crud protein; Ca: calcium; AP: available phosphate and DM: dry matter.

 

Table 5 Chemical analysis of protease enzyme

 

The highest EBI were observed by 20% replacing SBM with CSM without enzymes. As well as no significant response was found on relative weight of carcass, breast and thigh by treatments. The breast, thigh, wings, neck and back were not differences by experimental diets contain CSM levels (Elangovan et al. 2006). Since these organs were muscular and required to high quality protein for growth, in contrast protein digestibility the CSM is lower than soybean meal (80.9% vs. 91.67%).

 

Table 6 Effects of dietary cotton seed meal and protease enzyme on performance of broiler chickens

BWG: body weight gain; FI: feed intake; FCR: feed conversion ratio; EBI: European broiler index and CSM: cottonseed meal.

1: Control without enzyme; 2: Control with enzyme; 3: 20% replacing CSM with SBM without enzyme; 4: 20% replacing CSM with SBM with enzyme; 5: 30% replacing CSM without SBM without enzyme and 6: 30% replacing CSM with SBM without enzyme.

The means within the same row with at least one common letter, do not have significant difference (P>0.05).

SEM: standard error of the means.

 

Table 7 Effect of dietary cotton seed meal and protease enzyme on organs relative weight (% of live body weight) of broiler chicks

CSM: cottonseed meal.

1: Control without enzyme; 2: Control with enzyme; 3: 20% replacing CSM with SBM without enzyme; 4: 20% replacing CSM with SBM with enzyme; 5: 30% replacing CSM without SBM without enzyme and 6: 30% replacing CSM with SBM without enzyme.

The means within the same row with at least one common letter, do not have significant difference (P>0.05).

SEM: standard error of the means.

 

Table 8 Effect of dietary cotton seed meal and protease enzyme on protein and dry matter digestibility (%) and blood parameters

CSM: cottonseed meal.

1: Control without enzyme; 2: Control with enzyme; 3: 20% replacing CSM with SBM without enzyme; 4: 20% replacing CSM with SBM with enzyme; 5: 30% replacing CSM without SBM without enzyme and 6: 30% replacing CSM with SBM without enzyme.

The means within the same row with at least one common letter, do not have significant difference (P>0.05).

SEM: standard error of the means.

 

Table 9 Effect of dietary cotton seed meal and protease enzyme on jejunum morphology in broiler chickens at 42 days of age (μm)

CSM: cottonseed meal.

1: Control without enzyme; 2: Control with enzyme; 3: 20% replacing CSM with SBM without enzyme; 4: 20% replacing CSM with SBM with enzyme; 5: 30% replacing CSM without SBM without enzyme and 6: 30% replacing CSM with SBM without enzyme.

The means within the same row with at least one common letter, do not have significant difference (P>0.05).

SEM: standard error of the means.

 

Therefore no significant improved in breast and thigh growth were shown by CSM as lower protein source (Nie et al. 2015; Jazi et al. 2017). The tissues of the target organs (heart, liver and bursa of Fabricius) were also affected by treatments. In this study, proventriculus, pancreas and gizzard weight were significantly increased by CSM levels which may be due to high fiber content. Dietary structural components, such as fiber, have a attracted considerable attention as they may be increased large, well-developed and improved gizzard function (Kheravii et al. 2017). Therefore, gizzard was able to grind feed particles more completely and pancreatic enzyme secretion elevated by increased release of cholecystokinin (Amerah et al. 2007). Decreased dry matter in vitro digestibility, impairs and decreased the absorption of nutrients by anti-nutritional factors such as gossypol and cyclopropenoid fatty acids with higher levels of dietary CSM (Liu et al. 2006). The highest protein digestibility was observed by 30% replacing SBM with CSM with enzyme but no correspond with study of Cheng and Hardy, (2002). Also, some researches have noted that CSM reduced amino acid digestibility, this may be increased the demand of dietary synthetic amino acid supplementation in poultry performance. Mireles-Arriaga et al. (2015) have reported that ideal digestibility of protein increased by protease enzyme supplementation. Reduction plasma cholesterol by 30 and 20% SBM replacement with CSM in the diet may be due to its high fiber content (9.8% crude fiber). Reduces intestinal transit time and increased excretion of sterols (increased secretion of bile) by fiber binding with bile salt in the gastrointestinal tract. This could affects cholesterol metabolism and ultimately leads to the lower plasma cholesterol levels (Mateos et al. 2012; Taheri et al. 2016). In the study of Elangovan et al. (2006), cottonseed meal levels have no significant response on plasma protein which supported these results. No significant differences were observed in intestinal morphology by treatments. These results were in contrast with the results of some researches, which indicated fermented cottonseed meal increased villus height and villus height to crypt depth ratio.

 

CONCLUSION

In conclusion, no differences were found on performance by replacement of different levels of soybean meal with cottonseed meal in broiler chicken diet. Due to the low price of CSM up to 30% of cotton seed meal could be used in broiler chicken diet. Improved protein digestibility was shown by 30% replacement of SBM byCSMwith200 g/ton protease enzyme reaction.

 

ACKNOWLEDGEMENT

The authors thanks of Department of Animal Science of Bu-Ali Sins University with excellent scientific collaboration and financial support in this respect.

مراجع

Amerah A., Ravindran V., Lentle R. and Thomas D. (2007). Feed particle size: Implications on the digestion and performance of poultry. World's Poult. Sci. J. 63, 439-455.

AOAC. (2000). Official Methods of Analysis. 17^th Ed. Association of Official Analytical Chemists, Arlington, Washington, DC., USA.

Azman M.A. and Yilmaz M. (2005). The growth performance of broiler chicks fed with diets containing cottonseed meal supplemented with lysine. Rev. Méd. Vét. 156(2), 104-106.

Blevins S., Siegel P.B., Blodgett D.J., Ehrich M., Saunders G.K. and Lewis R.M. (2010). Effects of silymarin on gossypol toxicosis in divergent lines of chickens. Poult. Sci. 89, 1878-1886.

Cheng Z.J. and Hardy R.W. (2002). Apparent digestibility coefficients and nutritional value of cottonseed meal for rainbow trout (Oncorhynchus mykiss). Aquaculture. 212, 361-372.

Elangovan A., Tyagi P.K., Shrivastav A., Tyagi P.K. and Mandal A. (2006). GMO (Bt-Cry1Ac gene) cottonseed meal is similar to non-GMO low free gossypol cottonseed meal for growth performance of broiler chickens. Anim. Feed Sci. Technol. 129, 252-263.

Heidarinia A. and Malakian M. (2011). Nutritional evaluation of cottonseed meal with and without ferrous sulfate for broiler chickens. Res. J. Poult. Sci. 4, 14-17.

Jazi V., Boldaji F., Dastar B., Hashemi S.R. and Ashayerizadeh A. (2017). Effects of fermented cottonseed meal on the growth performance, gastrointestinal microflora population and small intestinal morphology in broiler chickens. British Poult. Sci. 58(4), 402-408.

Jin S.H., Corless A. and Sell J. (1998). Digestive system development in post-hatch poultry. World's Poult. Sci. J. 54, 335-345.

Kaczmarek S., Rogiewicz A., Mogielnicka M., Rutkowski A., Jones R.O. and Slominski B.A. (2014). The effect of protease, amylase, and nonstarch polysaccharide-degrading enzyme supplementation on nutrient utilization and growth performance of broiler chickens fed corn-soybean meal-based diets. Poult. Sci. 93, 1745-1753.

Kheravii S.K., Swick R.A., Choct M. and Wu S.B. (2017). Coarse particle inclusion and lignocellulose-rich fiber addition in feed benefit performance and health of broiler chickens. Poult. Sci. 96, 3272-3281.

Liu Y., Song G.L., Yi G.F., Hou Y., Huang J.W., Vazquez-Anon M. and Knight C. (2006). Effect of supplementing 2-hydroxy-4-(methylthio) butanoic acid and DL-methionine in corn-soybean-cottonseed meal diets on growth performance and carcass quality of broilers. Asian Australasian J. Anim. Sci. 19, 1197-1205.

Mateos G., Jiménez-Moreno E., Serrano M. and Lázaro R. (2012). Poultry response to high levels of dietary fiber sources varying in physical and chemical characteristics. J. Appl. Poult. Res. 21, 156-174.

Mireles-Arriaga A.I., EspinosaAyala E., Hemández-Garcia P.A. and Márquez-Molina O. (2015). Use of exogenous enzyme in animal feed. Life Sci. J. 12(2), 23-32.

Nagalakshmi D., Rao S.V.R., Panda A.K. and Sastry V.R. (2007). Cottonseed meal in poultry diets: A review. J. Poult. Sci. 44, 119-134.

Nazemzadeh S., Heshmat G. and Ansari H. (2017). Effect of supplemented ProAct (CT) Protease enzyme on performance and the amount of protein excreted in feces of broiler chickens. J. Livest. Sci. 8, 115-121.

Nie C., Zhang W., Ge W., Wang Y., Liu Y. and Liu J. (2015). Effects of fermented cottonseed meal on the growth performance, apparent digestibility, carcass traits, and meat composition in yellow-feathered broilers. Turkish J. Vet. Anim. Sci. 39, 350-356.

Parsons C. (2004). Gastrointestinal development and nutrient digestion in chicks. Pp. 169-176 in Proc. 25^th Western Nutr. Conf., Saskatoon, Canada.

SAS Institute. (2004). SAS^®/STAT Software, Release 9.4. SAS Institute, Inc., Cary, NC. USA.

Shawrang P., Mansouri M., Sadeghi A. and Ziaie F. (2011). Evaluation and comparison of gamma-and electron beam irradiation effects on total and free gossypol of cottonseed meal. Rad. Phys. Chem. 80, 761-762.

Taheri H., Tanha N. and Shahir M. (2016). Effect of wheat bran inclusion in barley-based diet on villus morphology of jejunum, serum cholesterol, abdominal fat and growth performance of broiler chickens. J. Livest. Sci. Technol. 4, 9-16.

Tang J., Sun H., Yao X.H., Wu Y.F., Wang X. and Feng J. (2012). Effects of replacement of soybean meal by fermented cottonseed meal on growth performance, serum biochemical parameters and immune function of yellow-feathered broilers. Asian-Australasian J. Anim. Sci. 25, 393-407.

Van Soest P.V., Robertson J. and Lewis B. (1991). Methods for dietary fiber, neutral detergent fiber, and nonstarch polysaccharides in relation to animal nutrition. J. Dairy Sci. 74, 3583-3597.

Xiong J., Wang Z.J., Miao L.H., Meng F.T. and Wu L.Y. (2016). Growth performance and toxic response of broilers fed diets containing unfermented or fermented cottonseed meal. J. Anim. Sci. 94, 457-457.