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Dietary Inclusion of Pancreatin Enzyme on the Ileal and Fecal Digestibility of Nutrients in Layer-Type Cockerels

ABSTRACT

The current experiment aimed to evaluate the effects of Pancreatin supplementation at different levels on ileal and fecal digestibility in layer-type cockerels. A total of 480-day-old silver brown Hy-Line male chicks were randomly allocated into 5 treatments, 6 replicates (16 birds per pen) arranged in a completely randomized design. Pancreatin enzyme was supplemented on a basal corn-soybean meal-based diet at 0, 250, 500, 750, and 1000 mg/kg and was fed in two growth phases (starter and grower). The results indicated that at the end of the starter stage, except for 1000 mg/kg, dietary Pancreatin supplementation levels increased (p<0.05) the ileal crude protein (CP). Similarly, addition of Pancreatin increased (p< 0.05) apparent ileal amino acids (AA) digestibility (AIAAD) total means of AA (MTAA), means of indispensable AA (MIAA) and dispensable AA (MDAA) with the optimum performance on 250 mg/kg and 500 mg/kg. However, except for histidine and alanine which were negatively affected (p<0.05), and MIAA, MDAA, MTAA which were also positively affected, the addition level at 1000 mg/kg did not affect most of the AIAAD compared to the non-supplemented. Further, Pancreatin supplementation had no effect (p>0.05) on nitrogen digestibility (ND), nitrogen retention (NR), digestible energy (DE), apparent metabolizable energy (AME), dry matter digestibility (DMD), dry matter retention (DMR), and apparent metabolizable energy corrected for nitrogen (AME-n) on fed starter diet. On the other hand, at the end of the grower stage, dietary Pancreatin enzyme supplementation reduced (p<0.05) the ileal CP, MIAA, MDAA, MTAA, AIAAD, AME, AME-n, ND, NR, DE, DMD, and DMR in a dose-dependent manner. The rate of reduction was more marked on Pancreatin addition level 1000 mg/kg. In conclusion, Pancreatin supplementation at 250 mg/kg, 500 mg/kg, and 750 mg/kg improved AIAAD and ileal CP, especially at the young age. The rate of pancreatin enzyme effect was dependent on enzyme supplement level to the ileal CP and individual amino acid.

Keywords:
Amino acid; Corn-SBM-based diet; Digestibility; Energy; Pancreatin

INTRODUCTION

Enzyme supplementation has been an innovative technology to maximize the growth of birds with increasing nutritional, environmental, and economical sustainability. Exogenous enzymes increase the growth of birds through their ability not only to reduce the adverse effects of anti-nutritional factors (non-starch polysaccharide) but also to decrease cell membrane integrity, changes the gut microflora which consequently improves utilization of protein solubility and energy digestibility (Olukosi et al., 2015Olukosi OA, Beeson LA, Englyst K, Romero LF. Effects of exogenous proteases without or with carbohydrases on nutrient digestibility and disappearance of non-starch polysaccharides in broiler chickens. Poultry Science 2015;94(11):2662-9.; Singh et al., 2017Singh AK, Berrocoso JFD, Dersjant-Li Y, Awati A, Jha R. Effect of a combination of xylanase, amylase and protease on growth performance of broilers fed low and high fiber diets. Animal Feed Science and Technology 2017;232:16-20.). Several commercially produced enzyme products composed of pure single digestive enzymes (protease, amylase, and lipase) have been used technically to improve nutrient digestibility and growth by augmenting the insufficient quantities of enzymes produced by the animal (Cowieson et al., 2019; Jabbar et al., 2021aJabbar A, Tahir M, Alhidary IA, Abdelrahman MA, Albadani H, Khan RU, et al. Impact of microbial protease enzyme and dietary crude protein levels on growth and nutrients digestibility in broilers over 15-28 days. Animals 2021a;11(9):2499.). Although, numerous studies have indicated that cocktails of these enzymes deliver more significant improvements in inherent digestibility, such as apparent metabolizable energy (AME), amino acids (AA) digestibility, nutrient availability, and to a lesser extent, reduced nutrient excretion, their result in corn soybean meal (SBM)-based diet has been inconsistent (Romero et al., 2013Romero LF, Parsons CM, Utterback PL, Plumstead PW, Ravindran V. Comparative effects of dietary carbohydrases without or with protease on the ileal digestibility of energy and amino acids and AMEn in young broilers. Animal Feed Science and Technology 2013;181(1/4):35-44.; 2014; Kaczmarek et al., 2014Kaczmarek SA, Rogiewicz A, Mogielnicka M, Rutkowski A, Jones RO, Slominski BA. 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. Poultry Science 2014;93(7):1745-53.; Adebiyi & Olukosi, 2015Adebiyi AO, Olukosi OA. Metabolizable energy content of wheat distillers' dried grains with solubles supplemented with or without a mixture of carbohydrases and protease for broilers and turkeys. Poultry Science 2015;94(6):1270-6.).

Pancreatin is a multiple-enzyme product usually extracted from animals’ pancreas. The enzyme has been used efficaciously with proven evidence in enzymatic deficiencies or replacement therapy in fish (Souza et al., 2020Souza APL, Ferreira TH, Mouriño JLP, Martins ML, Magenta Magalhaes AR, Tsuzuki MY. Use of Artemia supplemented with exogenous digestive enzymes as sole live food increased survival and growth during the larviculture of the longsnout seahorse Hippocampus reidi. Aquaculture Nutrition2020;26(3):964-77.), humans (Trang, 2014Trang T. Pancreatic enzyme replacement therapy for pancreatic exocrine insufficiency in the 21 st century. World Journal of Gastroenterology 2014;20(33):11467.), pigs (Rengman et al., 2010Rengman S, Fedkiv O, Botermans J, Svendsen J, Weström B, Pierzynowski S. The growth of exocrine pancreatic insufficient young pigs fed an elemental diet is dependent on enteral pancreatin supplementation. Livestock Science 2010;134(1-3):50-2.), cats and dogs (Xenoulis, 2020Xenoulis PG. Exocrine pancreatic insufficiency in dogs and cats. Clinical Small Animal Internal Medicine 2020;30(6):583-90.). In humans, this enzyme has been used for the treatment of pancreatic insufficiency with related conditions such as chronic pancreatitis, cystic fibrosis, and pancreatic carcinoma, which affect nutrient digestion (Trang 2014). Pancreatin enzyme is beneficial in mimicking its natural enzymatic properties in re-establishing the digestive conditions. Thus, pancreatin enzymes are used as a pivotal substitute for the physiological function of the pancreas to enhance digestion and absorption of nutrients passing through the intestines without being digested. Rengman et al. (2010) indi-cated that prepared oral pancreatin enzyme, with digestive properties, stimulated nutrient assimilation and anabolic processes in young fast-growing pigs. Similarly, the addition of pancreatin could improve the nutritional state by altering the intestinal microbiota in mice (Nishiyama et al., 2017).

Despite these hypothetical possibilities, Pancreatin enzyme has been one of the least studied in poultry nutrition even though it has been used commercially for more than two decades. Although studies on Pancreatin supplementation in pigs has been reported (Cervantes et al., 2011Cervantes M, Gómez R, Fierro S, Barrera MA, Morales A, Araiza BA, et al. Ileal digestibility of amino acids, phosphorus, phytate and energy in pigs fed sorghum-based diets supplemented with phytase and Pancreatin(r). Journal of Animal Physiology and Animal Nutrition 2011;95(2):179-86.), studies to determine appro-priate Pancreatin enzyme supplementation in the diet of poultry has not been reported. Accordingly, the present study was designed to determine the effect of dietary pancreatin supplementation on the ileal and fecal digestibility of nutrients in layer-type cockerels fed corn -SBM-based diet.

MATERIALS AND METHODS

Ethical statement

The current study was conducted in the research unit at the College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu Province. All experimental procedures and conducts were approved by the Care Advisory Committee of Yangzhou University Animal Ethic of Practice, Council of State, People Republic of China.

Experimental birds and Husbandry

A total of 480-day-old male chicks (Hy-Line Silver), with an average body weight (BW) 39.58 ±0.24 g from the Hy-line Company, were randomly assigned to 30-floor pens of 16 birds with wood shavings serving as litter material. Each pen with a floor space of 0.064 m2 was equipped with one round bottom plastic feeder and a round manual drinker. Before the birds’ arrival, disinfection was done with fumigation (mixer of formalin 40% with potassium permanganate powder). During the first week, the room temperature was maintained at 33 ºC and gradually decreased to suit the birds with adequate ventilation. Chicks were fed with fresh feed and water at ad libitum during the entire period. The birds were vaccinated against Marek’s disease at day old, Newcastle (Nobilis ND clone 30) on 14 d, and infectious bursal disease (Intervet) on 28 d by eye drop following manufacturers’ recommendations.

Diet formulations and Enzyme

The experimental diets typically of a corn-SBM-based diet was formulated at the Yangzhou University experimental feed mixing plan. The diets were fed in mash form for 70 days and in 2 phases based on growing period; Phase 1: Starter (1-42 d), and Phase 2: Grower (43-70 d). Treatment diets contained 11.98 MJ/kg of metabolizable energy (ME); 18.40% of crude protein (CP) at the starter phase and 11.85 MJ/kg of ME; 17.48 % of CP at the grower phase in a completely randomized design (Table 1). The Pancreatin enzyme extracted from pig pancreas was obtained from Shanghai Honest Biological Technology Co. Ltd. According to the supplier, the enzyme contained 561 U/g of protease, 3061 U/g of amylase, and 4352 U/g of lipase. The enzyme was added at 0, 250, 500, 750, and 1000 mg/kg, respectively, to the experimental diets. The dietary supplementation levels of the enzyme were considered based on the finding of Kim et al. (2018Kim J, Shim Y, Ingale SL, Hosseindoust A, Lee S, Rathi PC, et al. The microbial pH-stable exogenous multienzyme improved growth performance and intestinal morphology of weaned pigs fed a corn-soybean-based diet. Journal of Applied Animal Research 2018;46(1):559-65.) that enzyme supplementation from 250-1000 mg/kg in animal diets increased the total tract digestibility. Silicon oxide was incorporated at a level of 0.5% into the diets as an indigestible marker [for calculating acid insoluble ash (AIA)] at the last week of each feeding phase.

Table 1
Dietary formulations and nutrient concentration of the experimental diets (air-dry basis).

Experimental design and procedure

A week before the end of each phase, four birds with average weight were transferred into a wire flooring metabolic cage (70 cm2 per cage with 2 cm2 holes) equipped with a manual plastic drinker and a plastic feeder. Underneath each cage was a plastic tray for the excreta collection. According to the replicated diet per treatment, one metabolic cage was assigned to a floor pen’s unit cage. Fresh fecal samples were collected from each pen during the last 3 days of each feeding phase and immediately mixed, pooled by cage, and stored frozen at −20 °C for further analysis. At the end of each feeding phase (starter and growth), two birds per pen were selected and leg banded. Anesthetically, the birds were exposed to CO2 gas for approximately 30 s and euthanized before exsanguination. Ileal digesta was obtained from the intestine; thus, a section between Meckel’s diverticulum and ileo-cecal junction, about 2 cm in between, pooled per cage and immediately stored at -20 ºC for further analysis.

Sample collection and analyses

The experimental diets and fecal samples were air-dried in an oven at 60 ºC. The ileal digesta samples were also freeze-dried. The dried diets, fecal and ileal digesta were ground in a coffee grinder (CBG5 Smart Grind, Applica Consumer Products Inc., Shelton, CT, USA) through a 0.5 mm screen for analysis. The analysis of the dried ground samples is as follows; diet: ME, CP, Ash, AIA, Ca, P, C, AA. Ileal: AIA, AA, CP, DMD, DE, ND fecal:AIA DMR, NR, AME, AME-n. Samples were oven-dried at ±105 ºC for 16 hrs (method 934.01; AOAC, 2006) to determine the dry matter (DM) content. The CP content (N×6.25) was determined using the Kjeldahl method (method 984.13 A-D; AOAC, 2006) in an automated analyzer (Kjeltec 8400 Analyzer unit). Gross energy (GE) was determined in a bomb calorimeter (model 6200; Parr Instrument Co., Moline, IL). Samples for AA profiles were analyzed by HPLC (Hitachi L-8800 Amino Acid Analyzer, Tokyo, Japan) at University experimental laboratory followingmethod 982.30; AOAC (2006). The crude fiber determination method was done by sequential extraction with dilute acid and alkali (method 920.39; AOAC, 2006). The calcium (Ca) and phosphorus (P) analyses were as per method 975.03 B(b) and method 968.08, respectively, all of AOAC (2006). Silicon oxide concentration was determined using the technique described byDe Coca-Sinova et al. (2011).

Calculations

Ileal crude protein (CP) and amino acid digestibility (AIAAD) were calculated using acid insoluble acid (AIA) in the diets, digesta and excreta adopted from Ravindran et al. (1999Ravindran V, Hew LI, Ravindran G, Bryden WL. A comparison of ileal digesta and excreta analysis for the determination of amino acid digestibility in food ingredients for poultry. British Poultry Science 1999;40(2):266-74.):

A I A A D ( % ) = [ ( A A A I A ) i n d i e t ( A A A I A ) i n i l e a l ] ( A A A I A ) i n d i e t × 100 (1)

Apparent metabolizable energy (AME) was calculated according to Lammers et al. (2008Lammers PJ, Kerr BJ, Honeyman MS, Stalder K, Dozier WA, Weber TE, et al. Nitrogen-corrected apparent metabolizable energy value of crude glycerol for laying hens. Poultry Science 2008;87(1):104-7.):

D M R = ( A I A i n e x c r e t a A I A i n d i e t ) A I A i n e x c r e t a (2)

A M E ( M J k g ) = G E i n d i e t [ ( 1 D M R ) × G E i n e x c r e t a ] (3)

where DMR is the dry matter retention, AIA is the concentration of acid insoluble ash (g/kg), GE is the gross energy in the feed (MJ/kg)

Apparent metabolizable energy corrected for nitrogen (AME-n), dry matter digestibility (DMD) and digestible energy (DE)were also determined according to Yang et al. (2020Yang Z, Pirgozliev VR, Rose SP, Woods S, Yang HM, Wang ZY, Bedford MR. Effect of age on the relationship between metabolizable energy and digestible energy for broiler chickens. Poultry Science 2020;99(1):320-30.):

A M E n ( M J k g ) = G E i n d i e t ( G E i n e x c r e t a × A I A i n d i e t ) A I A i n e x c r e t a 34.39 × N r e t a i n e d 100 (4)

where DMR is the dry matter retention, AIA is the concentration of acid insoluble ash (%), GE is the gross energy (MJ/kg), 34.39 (MJ/kg) is the energy value of uric acid.

The N retained was calculated as:

N Re t a i n e d = N R i n d i e t [ N i n e x c r e t a × A I A i n d i e t ] A I A i n e x c r e t a (5)

where N is nitrogen (g/kg),thus birds per kilogram of diet consumed.

The nitrogen retention (NR) in excreta was calculated as described by Lammers et al. (2008Lammers PJ, Kerr BJ, Honeyman MS, Stalder K, Dozier WA, Weber TE, et al. Nitrogen-corrected apparent metabolizable energy value of crude glycerol for laying hens. Poultry Science 2008;87(1):104-7.):

N R = ( N i n d i e t A I A i n d i e t N i n e x c r e t a A I A i n e x c r e t a ) N i n d i e t A I A i n d i e t (6)

The digestible energy (DE) was calculated, using AIA as indigestible marker, as shown below:

D E ( M J k g ) = G E i n d i e t [ ( 1 D M R ) × G E i n e x c r e t a ] (7)

D M D = A I A i n i l e a l A I A i n d i e t A I A i n i l e a l (8)

where DMD is the dry matter digestibility, AIAis the concentration of acid insoluble ash (g/kg), and GEis the gross energy (MJ/kg).

The nitrogen digestibility (ND) was calculated as described by Lammers et al.(2008Lammers PJ, Kerr BJ, Honeyman MS, Stalder K, Dozier WA, Weber TE, et al. Nitrogen-corrected apparent metabolizable energy value of crude glycerol for laying hens. Poultry Science 2008;87(1):104-7.):

N D = N i n d i e t A I A i n d i e t N i n i l e a l A I A i n i l e a l N i n d i e t A I A i n d i e t (9)

where N represents nitrogen (g/kg) and AIA represents the concentration of AIA (g/kg).

Statistical analysis

The experimental data was initially processed by Microsoft Excel (Windows version 10) and were subjected to further analysis using ANOVA of SPSS 16.0 statistical software. Duncan’s multiple range tests were used to determine significant differences among treatment means. Unless specified otherwise, multiple comparisons with statistical differences were considered significant at p<0.05. The effect of increasing the enzyme-graded levels was variably assessed into linear and quadratic components using orthogonal polynomial contrasts for potential differences.

RESULTS

Growth performance

The present study’s growth performance data and discussion can be retrieved from Asare et al. (2021).

Ileal crude protein and amino acid digestibility

From table 2, data indicated that at the end of the starter phase, ileal CP digestibility was enhanced (p<0.05) by adding 250, 500, and 750 mg/kg of Pancreatin enzyme compared to the non-supplemented diet. Similarly, an increase (p<0.05) in AIAAD, total mean of AA (MTAA), and means of indispensable (MIAA) and dispensable AA (MDAA) was observed with dietary Pancreatin supplementation levels 250 mg/kg and 500 mg/kg. A linear increase (p<0.05) was observed on most of the AA under 250, and 500 mg/kg of Pancreatin enzyme except for histidine, leucine, phenylalanine, alanine, glutamine, proline, tyrosine and MDAA compared to the non-supplemented diet. Further, except for arginine, glutamine, and proline, 750 mg/kg of Pancreatin supplementation significantly increased (p<0.05) most of the ileal AA digestibility. In addition, with the exemption of histidine and alanine, which were negatively affected (p<0.05), and MIAA, MDAA, MTAA which was also positively affected (p<0.05), the addition level of 1000 mg/kg did not affect most of the AIAAD compared to the non-supplemented diet.

Table 2
Influence of Pancreatin supplements level on the coefficient of apparent ileal digestibility (%) of amino acids (AA) fed starter diet 1, 2.

On the other hand, at the end of the grower phase, dietary Pancreatin supplementation significantly decreased (p<0.05) the ileal CP digestibility, MIAA, MDAA, MTAA and AIAAD coefficients regardless of the supplementation level compared to the control diet. The rate of reduction was dependent on the type of amino acid to the Pancreatin supplementation level. However, a marked rate of reduction was observed on MIAA, MDAA, MTAA and AIAAD coefficients when Pancreatin enzyme was supplemented at 1000 mg/kg compared to the control diet. Similarly, a quadratic decrease was observed on histidine, proline and valine based on the level of Pancreatin supplementation (Table 3).

Table 3
Influence of Pancreatin supplements level on the coefficient of apparent ileal digestibility (%) of amino acids (AA) fed grower diet 1, 2.

Ileal and fecal nutrient determination

As shown in Table 4, there was no significant effect (p>0.05) on DMR, NR, AME, and AME-n when Pancreatin was supplemented on the starter diet. However, the addition of Pancreatin on a corn-SBM-based diet significantly lowered (p<0.05) the utilization of DMR, NR, AME, and AME-n compared to the non-supplemented diet on fed grower diet. Comparatively, the magnitude of reduction appeared to be more marked on 1000 mg/kg of Pancreatin enzyme supplementation.

Table 4
The effect of Pancreatin supplementation on fecal nutrient retention and metabolizable energy determination of cockerels1, 2.

Additionally, there was no significant effect (p>0.05) on ileal DMD, DE, and ND when Pancreatin was supplemented at different levels on the starter diet. However, irrespective of Pancreatin supplementation level on the grower diet, a decrease (p<0.05) in DMD, DE, and ND was observed compared to the control. A similar pattern of diminishing marginal response was observed quadratically (p=0.019) on the ND under the Pancreatin enzyme supplemented groups at the grower diet (Table 5).

Table 5
The effect of Pancreatin supplementation on ileal nutrient retention and digestible energy determination of birds1,2.

DISCUSSION

Increased digestible protein, carbohydrates, and other nutrients are necessary for the optimal growth in birds. Even though corn -SBM- based diet are suggested not viscous, the rapid passage of some nutrients, especially digestible protein in the gut, may significantly affect the deficiency of innate enzymes. Noy & Sklan (1995Noy Y, Sklan D. Digestion and absorption in the young chick. Poultry Science 1995;74(2):366-73.) and Jin et al. (1998Jin SH, Corless A, Sell JL. Digestive system development in post-hatch poultry. World's Poultry Science Journal 1998;54(4):335-45.) indicated that specific enzymes like lipase, amylase, and protease are needed to spearhead the digestion and absorption of nutrients. The present findings support literature findings indicating that pancreatin supplementation at the young stage played a double role, and except for its digestive properties, it increased the digestibility of the ileal CP and AIAAD (Rengman et al., 2010Rengman S, Fedkiv O, Botermans J, Svendsen J, Weström B, Pierzynowski S. The growth of exocrine pancreatic insufficient young pigs fed an elemental diet is dependent on enteral pancreatin supplementation. Livestock Science 2010;134(1-3):50-2.; Nishiyama et al., 2017). The pancreatin enzyme was markedly more effective at improving AIAAD, on a dose equivalent basis, and at a lowest dose of 250 mg/kg achieved a greater improvement then to 500 and 750 mg/kg. However, 1000 mg/kg had no effect on CP and several AA except for MIAA and MTAA which were increased at the young age compared to the non-supplement diet. In addition, a diminishing response was exerted with increased inclusion level 1000 mg/kg on histidine and alanine on fed supplemented starter diet. In some experiments, a marked increase in ileal digestibility of CP and AA was attributed to supplementing enzyme at 1000 mg/kg in bird’s diet. Published studies have shown that multi enzyme addition could be effective in improving the nutritive value of low- viscous, corn -SBM- based diet (Kim et al., 2018Kim J, Shim Y, Ingale SL, Hosseindoust A, Lee S, Rathi PC, et al. The microbial pH-stable exogenous multienzyme improved growth performance and intestinal morphology of weaned pigs fed a corn-soybean-based diet. Journal of Applied Animal Research 2018;46(1):559-65.; Saleh et al., 2020Saleh AA, Dawood MM, Badawi NA, Ebeid TA, Amber KA, Azzam MM. Effect of supplemental serine-protease from Bacillus licheniformis on growth performance and physiological change of broiler chickens. Journal of Applied Animal Research 2020;48(1):86-92.). The current results were consistent with these authors indicating that multi enzyme addition at 250 to 750 mg/kg added to birds’ diet has been efficient in enhancing the digestion of undigested protein passing through the gut. At the end of growth phase, adverse effects in ileal CP and AIAAD were found in response to Pancreatin supplementation at varied levels. Again, enzyme effects were dose-dependent, and at the highest pancreatin supplemented level (1000 mg/kg) compared to the control negatively affected the ileal CP digestibility, MIAA, MDAA, MTAA and AIAAD coefficients. Contrary to the present finding, Saleh et al. (2020) reported that AA and CP digestibility was increased by multi enzyme irrespective of the supplementation levels. Cowieson et al. (2019) indicated that although matured birds can secrete enough gastro-mucosal enzymes to enhance the consistent supply of digestible CP and AA stability, dietary supplementation with multi-enzyme complexes is noted to give high efficiency in improving nutrient digestibility in grow-finisher birds (Ravindran & Son 2011). The difference is probably due to the fact that the enzyme supplementation levels could have interfered with the digestive enzyme released and gut development to alter nutrient digestibility (Romero et al., 2014Romero LF, Sands JS, Indrakumar SE, Plumstead PW, Dalsgaard S, Ravindran V. Contribution of protein, starch, and fat to the apparent ileal digestible energy of corn- and wheat-based broiler diets in response to exogenous xylanase and amylase without or with protease. Poultry Science 2014;93(10):2501-13.; Amerah et al., 2017Amerah AM, Romero LF, Awati A, Ravindran V. Effect of exogenous xylanase, amylase, and protease as single or combined activities on nutrient digestibility and growth performance of broilers fed corn/soy diets. Poultry Science 2017;96(4):807-16.). In terms of the release of individual AA by supplemented Pancreatin enzyme, all the AA were readily released from corn- SBM -diet under supplementation levels 250 and 500 mg/kg on fed starter diet. Kim et al. (2018) have reported an increase in CP digestibility in response to increasing supplementation levels of multi-enzyme in fed corn-SBM-based diet. The current study agreed well with those reported earlier of corn-SBM- based diet for broiler chicken that the effect of adding enzymes to broilers diet fed at the starter phase is more significant compared to that during the growth phase (Mohammadigheisar & Kim, 2018Mohammadigheisar M, Kim IH. Addition of a protease to low crude protein density diets of broiler chickens. Journal of Applied Animal Research 2018;46(1):1377-81.). This suggests that birds at the starter phase might be more receptive to supplementary enzymes because of the enzymes ability to augment the insufficient endogenous enzymes synthesized by their immature gut (Uni et al., 1999Uni Z, Noy Y, Sklan D, Posthatch development of small intestinal function in the poult. Poultry Science 1999;78(2):215-22.; Dosković et al., 2013Dosković V, Bogosavljević-Bosković S, Pavlovski Z, Milošević B, Škrbić Z, Rakonjac S, et al. Enzymes in broiler diets with special reference to protease. World’s Poultry Science Journal 2013;69(2):343-60.). Therefore, one possible explanation for this difference in Pancreatin supplementation between the two growth phases could be attributed to bird’s age.

To our knowledge, the usage of either exogenous protease, amylase, or lipase enzymes as “standalone” has a direct positive contribution to energy digestibility and nutrient retention from the main energy-yielding substrates at the ileal and total tract level in birds (Liu et al., 2016Liu S, Feng L, Jiang WD, Liu Y, Jiang J, Wu P, et al. Impact of exogenous lipase supplementation on growth, intestinal function, mucosal immune and physical barrier, and related signaling molecules mRNA expression of young grass carp (Ctenopharyngodon idella). Fish & Shellfish Immunology 2016;31(12):88-105.; Amerah et al., 2017Amerah AM, Romero LF, Awati A, Ravindran V. Effect of exogenous xylanase, amylase, and protease as single or combined activities on nutrient digestibility and growth performance of broilers fed corn/soy diets. Poultry Science 2017;96(4):807-16.; Jabbar et al., 2021bJabbar A, Tahir M, Khan RU, Ahmad N. Interactive effect of exogenous protease enzyme and dietary crude protein levels on growth and digestibility indices in broiler chickens during the starter phase. Tropical Animal Health and Production 2021b;53(1):1- 3.). However, results have showed the synergistic and superior effect on the combination of these enzymes than protease, amylase, or lipase alone on the utilization of energy and nutrient retention. In the current study, Pancreatin supplementation did not affect the ileal DMD, DE, and ND or fecal DMR, NR, AME, and AME-n digestibility at the young age, irrespective of the supplementation level. Additionally, Pancreatin enzyme negatively affected both ileal DMD, DE, ND and fecal DMR, NR, AME, and AME-n digestibility at the grower stage irrespective of the supplementation level. It had been assumed that the use of Pancreatin enzyme would indirectly alter the gut microflora or bacterial population in the digestive system to ferment substantially the long-chain carbohydrate molecules used by some bacteria in the digestive tract as bird ages (Nishiyama et al. 2018Nishiyama H, Nagai T, Kudo M, Okazaki Y, Azuma Y, Watanabe T, et al. Supplementation of pancreatic digestive enzymes alters the composition of intestinal microbiota in mice. Biochemical and Biophysical Research Communications 2018;495(1):273-9.). However, the lack of effect on Pancreatin enzyme supplementation could be attributed to the fact that the enzyme levels could not disrupt the protein-starch interactions in the feed to short fragments digestible by innate pancreatic enzymes.

Contrary to the current work, Lee et al. (2018Lee JW, Patterson R, Woyengo TA. Porcine in vitro degradation and fermentation characteristics of canola co-products without or with fiber-degrading enzymes. Animal Feed Science and Technology 2018;241(3):133-40.) reported a positive effect on the digestibility of canola co-products and reduced fermentation of the resulting undigested nutrients by exogenous enzyme, porcine pepsin, and pancreatin in an in vitro experiment. However, the findings on total tract dry matter, starch digestibility and AME-n content of the current study agree with those of Cervantes et al. (2011Cervantes M, Gómez R, Fierro S, Barrera MA, Morales A, Araiza BA, et al. Ileal digestibility of amino acids, phosphorus, phytate and energy in pigs fed sorghum-based diets supplemented with phytase and Pancreatin(r). Journal of Animal Physiology and Animal Nutrition 2011;95(2):179-86.) that Pancreatin supplementation level could affect feed utilization on sorghum-SBM-based diets fed to growing pigs. Also, Kaczmarek et al. (2014Kaczmarek SA, Rogiewicz A, Mogielnicka M, Rutkowski A, Jones RO, Slominski BA. 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. Poultry Science 2014;93(7):1745-53.) observed a reduction in nutrient digestibility in enzyme supplemented corn-SBM-based diet due to endogenous pancreatic enzyme inhibition required for starch digestion. Although, several factors including feed processing, particle size, starch content in the feed, and non-feed-related factors such as bird’s genetics and age may account for the contradictory findings (Zaefarian et al., 2016Zaefarian F, Abdollahi MR, Ravindran V. Particle size and feed form in broiler diets:impact on gastrointestinal tract development and gut health. World's Poultry Science Journal 2016;72(2):277-90.). The reduction in nutrient retention and metabolizable energy digestibility at the grower phase which was dependent on the supplementation level in the current study could have also been influenced by the limited endogenous pancreatic enzyme secretions due to the supplementation of exogenous enzymes (Kaczmarek et al., 2014).

CONCLUSION

The present findings indicated that Pancreatin supplementation at 250-750 mg/kg could enhance the ileal crude protein and amino acid digestibility of cockerels at a young age. The magnitude of increase depended on the enzyme supplement level to the individual amino acid. The current study suggested that Pancreatin enzyme can be an effective tool in poultry production, especially at the young age, to improve amino acid digestibility. However, there is a need for further research to determine the effects of Pancreatin enzyme supplementation in broiler production.

REFERENCES

  • Adebiyi AO, Olukosi OA. Metabolizable energy content of wheat distillers' dried grains with solubles supplemented with or without a mixture of carbohydrases and protease for broilers and turkeys. Poultry Science 2015;94(6):1270-6.
  • Amerah AM, Romero LF, Awati A, Ravindran V. Effect of exogenous xylanase, amylase, and protease as single or combined activities on nutrient digestibility and growth performance of broilers fed corn/soy diets. Poultry Science 2017;96(4):807-16.
  • AOAC - Association of official analytical chemists. Official methods of analysis. 18th ed. Arlington; 2006.
  • Asare E, Yang Z, Yang H, Wang Z. Evaluation of dietary Pancreatin as an exogenous enzyme on growth performance, gene expression, immunological responses, serum immunoglobins, and intestinal morphology in cockerels. Journal of Applied Animal Research 2022;50(1):61-8.
  • Cervantes M, Gómez R, Fierro S, Barrera MA, Morales A, Araiza BA, et al. Ileal digestibility of amino acids, phosphorus, phytate and energy in pigs fed sorghum-based diets supplemented with phytase and Pancreatin(r). Journal of Animal Physiology and Animal Nutrition 2011;95(2):179-86.
  • Coca-Sinova A de, Mateos GG, González-Alvarado JM, Centeno C, Garcia RL, Moreno EJ. Comparative study of two analytical procedures for the determination of acid insoluble ash for evaluation of nutrient retention in broilers. Spanish Journal of Agricultural Research 2011;9(3):761-8.
  • Dosković V, Bogosavljević-Bosković S, Pavlovski Z, Milošević B, Škrbić Z, Rakonjac S, et al. Enzymes in broiler diets with special reference to protease. World’s Poultry Science Journal 2013;69(2):343-60.
  • Hussein EOS, Suliman GM, Alowaimer AN, Ahmed SH, Abd El-Hack ME, Taha AE, et al. Growth, carcass characteristics, and meat quality of broilers fed a low-energy diet supplemented with a multienzyme preparation. Poultry Science 2020;99(4):1988-94.
  • Jabbar A, Tahir M, Alhidary IA, Abdelrahman MA, Albadani H, Khan RU, et al. Impact of microbial protease enzyme and dietary crude protein levels on growth and nutrients digestibility in broilers over 15-28 days. Animals 2021a;11(9):2499.
  • Jabbar A, Tahir M, Khan RU, Ahmad N. Interactive effect of exogenous protease enzyme and dietary crude protein levels on growth and digestibility indices in broiler chickens during the starter phase. Tropical Animal Health and Production 2021b;53(1):1- 3.
  • Jin SH, Corless A, Sell JL. Digestive system development in post-hatch poultry. World's Poultry Science Journal 1998;54(4):335-45.
  • Kaczmarek SA, Rogiewicz A, Mogielnicka M, Rutkowski A, Jones RO, Slominski BA. 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. Poultry Science 2014;93(7):1745-53.
  • Kim J, Shim Y, Ingale SL, Hosseindoust A, Lee S, Rathi PC, et al. The microbial pH-stable exogenous multienzyme improved growth performance and intestinal morphology of weaned pigs fed a corn-soybean-based diet. Journal of Applied Animal Research 2018;46(1):559-65.
  • Lammers PJ, Kerr BJ, Honeyman MS, Stalder K, Dozier WA, Weber TE, et al. Nitrogen-corrected apparent metabolizable energy value of crude glycerol for laying hens. Poultry Science 2008;87(1):104-7.
  • Lee JW, Patterson R, Woyengo TA. Porcine in vitro degradation and fermentation characteristics of canola co-products without or with fiber-degrading enzymes. Animal Feed Science and Technology 2018;241(3):133-40.
  • Liu S, Feng L, Jiang WD, Liu Y, Jiang J, Wu P, et al. Impact of exogenous lipase supplementation on growth, intestinal function, mucosal immune and physical barrier, and related signaling molecules mRNA expression of young grass carp (Ctenopharyngodon idella). Fish & Shellfish Immunology 2016;31(12):88-105.
  • Mohammadigheisar M, Kim IH. Addition of a protease to low crude protein density diets of broiler chickens. Journal of Applied Animal Research 2018;46(1):1377-81.
  • Nishiyama H, Nagai T, Kudo M, Okazaki Y, Azuma Y, Watanabe T, et al. Supplementation of pancreatic digestive enzymes alters the composition of intestinal microbiota in mice. Biochemical and Biophysical Research Communications 2018;495(1):273-9.
  • Noy Y, Sklan D. Digestion and absorption in the young chick. Poultry Science 1995;74(2):366-73.
  • Olukosi OA, Beeson LA, Englyst K, Romero LF. Effects of exogenous proteases without or with carbohydrases on nutrient digestibility and disappearance of non-starch polysaccharides in broiler chickens. Poultry Science 2015;94(11):2662-9.
  • Ravindran V, Hew LI, Ravindran G, Bryden WL. A comparison of ileal digesta and excreta analysis for the determination of amino acid digestibility in food ingredients for poultry. British Poultry Science 1999;40(2):266-74.
  • Rengman S, Fedkiv O, Botermans J, Svendsen J, Weström B, Pierzynowski S. The growth of exocrine pancreatic insufficient young pigs fed an elemental diet is dependent on enteral pancreatin supplementation. Livestock Science 2010;134(1-3):50-2.
  • Romero LF, Parsons CM, Utterback PL, Plumstead PW, Ravindran V. Comparative effects of dietary carbohydrases without or with protease on the ileal digestibility of energy and amino acids and AMEn in young broilers. Animal Feed Science and Technology 2013;181(1/4):35-44.
  • Romero LF, Sands JS, Indrakumar SE, Plumstead PW, Dalsgaard S, Ravindran V. Contribution of protein, starch, and fat to the apparent ileal digestible energy of corn- and wheat-based broiler diets in response to exogenous xylanase and amylase without or with protease. Poultry Science 2014;93(10):2501-13.
  • Saleh AA, Dawood MM, Badawi NA, Ebeid TA, Amber KA, Azzam MM. Effect of supplemental serine-protease from Bacillus licheniformis on growth performance and physiological change of broiler chickens. Journal of Applied Animal Research 2020;48(1):86-92.
  • Singh AK, Berrocoso JFD, Dersjant-Li Y, Awati A, Jha R. Effect of a combination of xylanase, amylase and protease on growth performance of broilers fed low and high fiber diets. Animal Feed Science and Technology 2017;232:16-20.
  • Souza APL, Ferreira TH, Mouriño JLP, Martins ML, Magenta Magalhaes AR, Tsuzuki MY. Use of Artemia supplemented with exogenous digestive enzymes as sole live food increased survival and growth during the larviculture of the longsnout seahorse Hippocampus reidi. Aquaculture Nutrition2020;26(3):964-77.
  • Trang T. Pancreatic enzyme replacement therapy for pancreatic exocrine insufficiency in the 21 st century. World Journal of Gastroenterology 2014;20(33):11467.
  • Uni Z, Noy Y, Sklan D, Posthatch development of small intestinal function in the poult. Poultry Science 1999;78(2):215-22.
  • Xenoulis PG. Exocrine pancreatic insufficiency in dogs and cats. Clinical Small Animal Internal Medicine 2020;30(6):583-90.
  • Yang Z, Pirgozliev VR, Rose SP, Woods S, Yang HM, Wang ZY, Bedford MR. Effect of age on the relationship between metabolizable energy and digestible energy for broiler chickens. Poultry Science 2020;99(1):320-30.
  • Zaefarian F, Abdollahi MR, Ravindran V. Particle size and feed form in broiler diets:impact on gastrointestinal tract development and gut health. World's Poultry Science Journal 2016;72(2):277-90.

Publication Dates

  • Publication in this collection
    09 Dec 2022
  • Date of issue
    2023

History

  • Received
    08 May 2022
  • Accepted
    28 July 2022
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