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Evaluation of liquid xylanase and phytase added after broiler feed pelletization

ABSTRACT

The objective of this study was to evaluate whether the addition of the enzymes phytase and xylanase, isolated or associated, in the liquid form after feed pelletization could improve energy utilization and digestibility of calcium and phosphorus by broiler chickens. Three experiments were performed using 120 birds each, divided into five treatments with eight replicates per group (n = 3), identified as: experiment 1 (xylanase: control, 1000 IU, 1500 IU, 2000 IU, 2500 IU), experiment 2 (phytase: control, 500 FTU, 1000 FTU, 1500 FTU, 2000 FTU), experiment 3 (xylanase + phytase: control, 3000 IU + 500 FTU, 3000 IU + 1000 FTU, 3000 IU + 1500 FTU, 3000 IU + 2000 FTU). Samples for digestibility tests were collected at 14 to 21 days of age. Therefore, the inclusion of liquid phytase and liquid phytase + xylanase after pelletization in broiler diets has become a relevant way to reduce the inclusion of inorganic P, which can reduce the cost of feed and P excretion in the environment. Furthermore, it is an interesting strategy to avoid enzyme denaturation in the pelleting process.

digestibility; exogenous enzymes; minerals; pelletizing

1. Introduction

Poultry diets are mainly based on corn, wheat, and soybean meal, which contain around 10 to 22.7% of non-starch polysaccharides (NSP) in their composition (Kim et al., 2021Kim, M.; Ingale, S. L.; Hosseindoust, A.; Choi, Y.; Kim, K. and Chae, B. 2021. Synergistic effect of exogenous multi-enzyme and phytase on growth performance, nutrients digestibility, blood metabolites, intestinal microflora and morphology in broilers fed corn-wheat-soybean meal diets. Animal Bioscience 34:1365-1374. https://doi.org/10.5713/ab.20.0663
https://doi.org/10.5713/ab.20.0663...
), classified as antinutritional factors that negatively impact the gastrointestinal tract of birds. Non-starch polysaccharides may increase the viscosity of the digesta, decrease the rate of intestinal passage of nutrients, interfere with the microbiota that colonizes the digestive tract (Cardoso et al., 2018Cardoso, V.; Fernandes, E. A.; Santos, H. M. M.; Maçãs, B.; Lordelo, M. M.; Telo Da Gama, L.; Ferreira, L. M. A.; Fontes, C. M. G. A. and Ribeiro, T. 2018. Variation in levels of non-starch polysaccharides and endogenous endo-1,4-β-xylanases affects the nutritive value of wheat for poultry. British Poultry Science 59:218-226. https://doi.org/10.1080/00071668.2018.1423674
https://doi.org/10.1080/00071668.2018.14...
), as well as increase the rate of mucosal cell turn-over, mucin secretion, and undigested content (Dong et al., 2018Dong, B.; Liu, S.; Wang, C. and Cao, Y. 2018. Effects of xylanase supplementation to wheat-based diets on growth performance, nutrient digestibility and gut microbes in weanling pigs. Asian-Australasian Journal of Animal Sciences 31:1491-1499. https://doi.org/10.5713/ajas.17.0867
https://doi.org/10.5713/ajas.17.0867...
). Therefore, they negatively influence poultry performance and health.

Birds do not produce endogenous enzymes to break beta 1,4 and alpha 1,6 bonds of some components present in vegetables. In this way, several exogenous enzymes such as xylanases, alpha-amylases, beta-glucanases, cellulases, and phytases are produced to aid in the digestion of nutrients, increase the proportion of beneficial bacteria in the intestinal tract, decrease intestinal viscosity, and save energy. These enzymes might be produced by some bacteria, yeasts, and fungi (Oliveira and Moraes, 2007Oliveira, M. C. and Moraes, V. M. B. 2007. Mananoligossacarídeos e enzimas em dietas à base de milho e farelo de soja para aves. Ciência Animal Brasileira 8:339-357. https://www.revistas.ufg.br/vet/article/view/1673
https://www.revistas.ufg.br/vet/article/...
). The addition of exogenous enzymes to feed can reduce the harmful effects caused by antinutritional factors such as NSP and phytic acid. It can also improve the digestive process and availability of energy in food, particularly in young birds (Olukosi et al., 2020Olukosi, O. A.; González-Ortiz, G.; Whitfield, H. and Bedford, M. R. 2020. Comparative aspects of phytase and xylanase effects on performance, mineral digestibility, and ileal phytate degradation in broilers and turkeys. Poultry Science 99:1528-1539. https://doi.org/10.1016/j.psj.2019.11.018
https://doi.org/10.1016/j.psj.2019.11.01...
).

In this context, phytase is the main exogenous enzyme used in the diet of birds, since they do not produce endogenous phytase (Dessimoni et al., 2019Dessimoni, G. V.; Sakomura, N. K.; Donato, D. C. Z.; Goldflus, F.; Ferreira, N. T. and Dalólio, F. S. 2019. Effect of supplementation with Escherichia coli phytase for broilers on performance, nutrient digestibility, minerals in the tibia and diet cost. Semina: Ciências Agrárias 40:767-780. https://doi.org/10.5433/1679-0359.2019v40n2p767
https://doi.org/10.5433/1679-0359.2019v4...
). Phytase breaks down phosphate from phytic acid in a sequential process, and the final product is myoinositol and phosphate, besides other nutrients that are trapped in phytic acid (Yu et al., 2012Yu, S.; Cowieson, A.; Gilbert, C.; Plumstead, P. and Dalsgaard, S. 2012. Interactions of phytate and myo-inositol phosphate esters (IP 1–5 ) including IP 5 isomers with dietary protein and iron and inhibition of pepsin. Journal of Animal Science 90:1824-1832. https://doi.org/10.2527/jas.2011-3866
https://doi.org/10.2527/jas.2011-3866...
). Kim et al. (2021)Kim, M.; Ingale, S. L.; Hosseindoust, A.; Choi, Y.; Kim, K. and Chae, B. 2021. Synergistic effect of exogenous multi-enzyme and phytase on growth performance, nutrients digestibility, blood metabolites, intestinal microflora and morphology in broilers fed corn-wheat-soybean meal diets. Animal Bioscience 34:1365-1374. https://doi.org/10.5713/ab.20.0663
https://doi.org/10.5713/ab.20.0663...
reported that the addition of multiple enzymes with phytase improves the digestibility of dry matter (DM), crude energy, crude protein, calcium, and phosphorus. Another advantage of utilizing phytase is that it increases the availability of phosphorus, reducing the use of inorganic phosphate (Dessimoni et al., 2019Dessimoni, G. V.; Sakomura, N. K.; Donato, D. C. Z.; Goldflus, F.; Ferreira, N. T. and Dalólio, F. S. 2019. Effect of supplementation with Escherichia coli phytase for broilers on performance, nutrient digestibility, minerals in the tibia and diet cost. Semina: Ciências Agrárias 40:767-780. https://doi.org/10.5433/1679-0359.2019v40n2p767
https://doi.org/10.5433/1679-0359.2019v4...
).

In addition, one can mention xylanases, which are carbohydrases that act on the xylan skeleton, breaking the internal beta-(1,4) bonds, thus reaching the cell wall (Zhang et al., 2016Zhang, Z.; Smith, C.; Li, W. and Ashworth, J. 2016. Characterization of nitric oxide modulatory activities of alkaline-extracted and enzymatic-modified arabinoxylans from corn bran in cultured human monocytes. Journal Agricultural Food Chemistry 64:8128-8137. https://doi.org/10.1021/acs.jafc.6b02896
https://doi.org/10.1021/acs.jafc.6b02896...
). In this way, they release oligosaccharides, disaccharides, and monomeric pentose sugars such as xylose. Then, xylose can be absorbed in the small intestine and provide energy (Huntley and Patience, 2018Huntley, N. F. and Patience, J. F. 2018. Xylose metabolism in the pig. Plos One 13:e0205913. https://doi.org/10.1371/journal.pone.0205913
https://doi.org/10.1371/journal.pone.020...
). Therefore, the use of xylanase in poultry diets can reduce digesta viscosity and improve nutrient availability by NSP hydrolysis (Barasch and Grimes, 2021Barasch, I. B. and Grimes, J. L. 2021. The effect of a heat-stable xylanase on digesta viscosity, apparent metabolizable energy and growth performance of broiler chicks fed a wheat-based diet. Poultry Science 100:101275. https://doi.org/10.1016/j.psj.2021.101275
https://doi.org/10.1016/j.psj.2021.10127...
), in addition to aiding the phytase access to phytate stored in the cell wall membrane of vegetables, which also helps to improve the use of phosphorus and energy.

Broiler feed is usually pelletized, a process by which the ingredients are subjected to temperatures between 75 and 83 ℃. Due to their thermolabile properties, these conditions may result in denaturation and inactivation of exogenous enzymes when subjected to pressure and heat (Pope et al., 2020Pope, J. T.; Brake, J. and Fahrenholz, A. C. 2020. Parameters monitored during the pelleting process and their relationship to xylanase activity loss. Animal Feed Science and Technology 259:114344. https://doi.org/10.1016/j.anifeedsci.2019.114344
https://doi.org/10.1016/j.anifeedsci.201...
). Given this fact, the application of enzymes after pelletization, in liquid form, might be a promising alternative to overcome this problem. Then, in a spraying process in the already pelleted feed, the enzymes can be added at room temperature without loss due to enzymatic denaturation that could occur at higher temperatures. Therefore, the objective of this study was to evaluate whether the addition of phytase and xylanase enzymes, isolated or associated, in liquid form after pelleting in broiler ration, could improve energy utilization and digestibility of calcium and phosphorus in the diets.

2. Material and Methods

2.1. Animals, experimental design, and diets

The experimental protocol was approved by the Institutional Ethics Committee on the Use of Animals (CEUA) under protocol number 27/2020.

Three experiments were performed with 360 Cobb 500 females and broiler chickens in Santa Catarina, Brazil (Latitude: −26.8364531149; Longitude: −52.4079407059). In the three trials, a completely randomized experimental design was adopted with five treatments (Table 1), eight replicates, and three animals per experimental unit. In the pre-experimental period (1 to 14 days), the birds were raised in shavings, according to the lineage manual.

Table 1
Treatments used – final levels in the feed

The basal ration used in this study was based on crushed corn and soybean meal according to the nutritional requirements described in the Brazilian Tables for Poultry and Swine (Rostagno et al., 2017Rostagno, H. S.; Albino, L. F. T.; Hannas, M. I.; Donzele, J. L.; Sakomura, N. K.; Perazzo, F. G.; Saraiva, A.; Teixeira, M. L.; Rodrigues, P. B.; Oliveira, R. F.; Barreto, S. L. T. and Brito, C. O. 2017. Tabelas brasileiras para aves e suínos: composição de alimentos e exigências nutricionais. 4.ed. Departamento de Zootecnia/UFV, Viçosa, MG.; Table 2), however, with a reduction in metabolizable energy levels (3150 to 3100 kcal/kg) and crude protein (19.71 to 19.00%). The rations were pelleted in a pelletizer model Koppers Junior C40 (Siemens), with a 50 horsepower (hp) engine and a ring with holes with a diameter of 3/16 inches, being processed at a temperature of 80 ℃ and device retention time of 30 s. The enzymes were added to the feed in liquid form, immediately after pelleting, with the aid of in-line sprinklers. The xylanase used had a concentration of 3,000 IU/g, and the phytase, a concentration of 10,000 FTU/g. Feed and water were provided ad libitum throughout the experimental period.

Table 2
Feed and nutritional composition of the basal experimental ration

2.2. Sampling and feed energy calculation

The total excreta collection method was used, with the birds housed in a metallic structure battery consisting of cages, distributed on two floors, equipped with a feeder and a nipple-type drinker. The experimental period was of ten days and consisted of five days for adapting the birds to cages, feed, and management, and five days for total excreta collection. Excreta from all experimental units were collected daily (08:00 and 17:00 h) in plastic-covered trays and conditioned in a freezer until the end of the experiment. At the end of the experimental period, the excreta were thawed, weighed, and homogenized, after which a 500 g sample was taken from each experimental unit and placed in a forced-circulation oven at 55 ℃ for pre-drying. They were analyzed according to the techniques described by Silva and Queiroz (2002)Silva, D. J. and Queiroz, A. C. 2002. Análise de alimentos: métodos químicos e biológicos. 3.ed. Universidade Federal de Viçosa, Viçosa, MG to determine the values of metabolizable energy (Matterson et al., 1965Matterson, L. D.; Potter, L. M.; Stutz, M. W. and Singsen, E. P. 1965. The metabolizable energy of feed ingredients for chickens. University of Connecticut, Agricultural Experiment Station, Storrs. Research Report, 7. 11p.) and nitrogen (Kjieldahl’s method), for subsequent use, obtaining the metabolizable energy values of the diets.

At the end of the experiment, the feed intake per experimental unit during the five days of the collection was determined. Once the results of the laboratory analyses of the rations and excreta were obtained, the apparent metabolizable energy (AME) and apparent metabolizable energy corrected by nitrogen retention (AMENR) values were calculated using equations proposed by Matterson et al. (1965)Matterson, L. D.; Potter, L. M.; Stutz, M. W. and Singsen, E. P. 1965. The metabolizable energy of feed ingredients for chickens. University of Connecticut, Agricultural Experiment Station, Storrs. Research Report, 7. 11p.:

Feed AME ( k c a l / k g ) = GE ingested GE excreted DM ingested (Eq.1)
Feed AMENR ( k c a l / k g ) = GE ingested ( GE excreted 8.22 × N B ) DM ingested (Eq.2)

in which GE = gross energy and NB = nitrogen balance (N ingested − N excreted).

2.3. Dietary calcium and phosphorus balance calculation

To determine the utilization of calcium and phosphorus in the diets, excreta and feed samples were subjected to absorption spectrophotometry analysis as described by Tedesco et al. (1995)Tedesco, M. J.; Gianello, C.; Bissani, C. A.; Bohnen, H. and Volkweiss, S. J. 1995. Análises de solo, plantas e outros materiais. 2.ed. Universidade Federal do Rio Grande do Sul, Porto Alegre. 174p.. From these data, the phosphorus digestibility coefficients were calculated according to the formula below:

Digestibility coefficient ( % ) = % of nutrient in feces % of nutrient in the diet × 100 (Eq.3)

2.4. Statistical analysis

The results were subjected to the Shapiro-Wilk normality test. When normal, the analysis of variance used was parametric and if significant, subjected to linear and quadratic regression analysis and also to Dunnett’s test. However, when there was no normality in the data, the variables were analyzed using the non-parametric Kruskal-Wails test and Dunnett’s test to compare means. The different levels of the exogenous enzymes were subjected to regression analysis using linear and quadratic functions. All analyzes were performed at a 5% significance level, using the R statistical program (R Development Core Team, 2019R Development Core Team. 2019. R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna.). The statistical model was as follows:

Y i j = μ + β i + ε i j , (Eq.4)

in which Yij = dependent variable, μ = variable mean, βi = fixed effect of broilers of the group, and εij = experimental error associated with observation Yij.

3. Results

There was no statistical difference for the variables DM intake, DM excreta production, AME, and AMENR (P>0.05) for broilers supplemented with different levels of xylanase in liquid form after pelleting (Table 3).

Table 3
Metabolizable energy values of diets supplemented with different levels of liquid xylanase after pelleting

There was no statistical difference for calcium balance variables for birds supplemented with different levels of phytase in liquid form (P>0.05; Table 4). However, for the phosphorus balance, a linear effect was observed for DM intake as the levels of phytase inclusion increased (P = 0.01). We also observed an increase in the total phosphorus ingested in the 1000 FTU group, followed by 2000 FTU group in relation to the 0 and 1500 FTU groups (P<0.001). In addition, there was this variable presented a linear (Y=0.3569x+5.4174) and also a quadratic effect (Y=0.3202x2+2.2781x+3.176). For the apparent digestibility coefficient (ADCo) of the total phosphorus of the diet, an increase was observed in all groups that received liquid phytase in relation to the control (P = 0.020), and there was also it also presented a linear (Y=2.5486x+60.347) and quadratic effect (Y=1.822x2+13.481x+47.593). All groups that received liquid phytase had lower phosphorus excretion compared with the control (P = 0.020), besides the linear (Y=2.5152x+39.506) and quadratic effect (Y=1.944x213.282x+52.067) (Table 5).

Table 4
Mean values of calcium balance of broiler rations supplemented with liquid phytase at different levels after pelleting
Table 5
Mean values of phosphorus balance in broiler rations supplemented with liquid phytase at different levels after pelleting

There was no statistical difference in the calcium balance of birds supplemented with different levels of liquid phytase + xylanase (P>0.05; Table 6). However, for the phosphorus balance, a decrease in DM intake was verified in birds that received 1000 FTU + 3000 IU in relation to the other groups (P<0.001). The largest amount of phosphorus ingested was observed in groups of 1500 and 2000 FTU, followed by groups 0 and 500 in relation to the group that received 1000 FTU of phytase + xylanase (P<0.001). The highest coefficient of total apparent digestibility of phosphorus was found in all groups that received liquid enzymes compared with the control (P = 0.046). Along with that, the lowest total phosphorus excretion was also observed in all groups that received the two enzymes associated with the control (P = 0.046; Table 7).

Table 6
Mean values of calcium balance of broiler rations supplemented with liquid xylanase + phytase at different levels after pelleting
Table 7
Phosphorus balance (mean values) in broiler rations supplemented with liquid xylanase + phytase at different levels after pelleting

4. Discussion

The addition of liquid phytase (500 FTU) and phytase + xylanase (500 FTU + 3000 IU) after pelleting showed promising values in terms of improving the use of phosphorus in the feed and, with that, decreasing phosphorus excretion, which is desirable in production systems due to the environmental impact that this mineral may cause. Furthermore, there are not many studies in the literature on the use of liquid enzymes after pelleting and the impact of this process on the denaturation of exogenous enzymes used in animal production. However, it is known that the factor that most contributes to the denaturation of enzymes is the accumulation of forces necessary to bind the particles within the pelletizing matrix, such as moisture, heat, and pressure, in addition to exposure to steam inside the conditioning chamber (Pope and Fahrenholz, 2020Pope, J. T. and Fahrenholz, A. C. 2020. The effect of the level of mixer-added water and mash conditioning temperature on parameters monitored during pelleting and phytase and xylanase thermostability. Animal Feed Science and Technology 269:114679. https://doi.org/10.1016/j.anifeedsci.2020.114679
https://doi.org/10.1016/j.anifeedsci.202...
).

In this context, when adding liquid xylanase after pelleting, no improvement in energy coefficients was observed. The results of the present study corroborate the results found by Denstadli et al. (2010)Denstadli, V.; Westereng, B.; Biniyam, H. G.; Balance, S.; Knutsen, S. H. and Svihus, B. 2010. Effects of structure and xylanase treatment of brewers’ spent grain on performance and nutrient availability in broiler chickens. British Poultry Science 51:419-426. https://doi.org/10.1080/00071668.2010.495745
https://doi.org/10.1080/00071668.2010.49...
, who pointed out that when supplementing broilers with 2500 IU/kg of xylanase in the liquid form after pelleting, there was no significant difference in AME. Thus, the inclusion levels used in this study were not efficient to cause the hydrolysis of the beta 1,4 and alpha 1,6 bonds of the NSP, so that there were significant changes in the metabolizable energy of the diet or there was not enough substrate for the enzyme to act efficiently. However, more studies should be carried out using different xylanases, as several authors demonstrate that the pelletization process decreases the activity of xylanases (Pope et al., 2020Pope, J. T.; Brake, J. and Fahrenholz, A. C. 2020. Parameters monitored during the pelleting process and their relationship to xylanase activity loss. Animal Feed Science and Technology 259:114344. https://doi.org/10.1016/j.anifeedsci.2019.114344
https://doi.org/10.1016/j.anifeedsci.201...
; Pope and Fahrenholz, 2020Pope, J. T. and Fahrenholz, A. C. 2020. The effect of the level of mixer-added water and mash conditioning temperature on parameters monitored during pelleting and phytase and xylanase thermostability. Animal Feed Science and Technology 269:114679. https://doi.org/10.1016/j.anifeedsci.2020.114679
https://doi.org/10.1016/j.anifeedsci.202...
).

The use of phytase improved the apparent digestibility coefficient of P and decreased its excretion. Sens et al. (2021)Sens, R. F.; Bassi, L. S.; Almeida, L. M.; Rosso, D. F.; Teixeira, L. V. and Maiorka, A. 2021. Effect of different doses of phytase and protein content of soybean meal on growth performance, nutrient digestibility, and bone characteristics of broilers. Poultry Science 100:100917. https://doi.org/10.1016/j.psj.2020.12.015
https://doi.org/10.1016/j.psj.2020.12.01...
found that the inclusion of 2500 FTU/kg phytase for broiler chickens increased phosphorus digestibility, plasma myoinositol levels, and tibia and toe ashes. Thus, by increasing the digestibility of this mineral, its excretion was reduced, and bone mineral composition was improved. Auereli et al. (2011)Auereli, R.; Faruk, M. U.; Cechova, I.; Pedersen, P. B.; Elvig-Joegensen, S. G.; Fru, F. and Broz, J. 2011. The efficacy of a novel microbial 6-phytase expressed in Aspergillus oryzae on the performance and phosphorus utilization in broiler chickens. International Journal of Poultry Science 10:160-168. https://doi.org/10.3923/ijps.2011.160.168
https://doi.org/10.3923/ijps.2011.160.16...
observed an increase in phosphorus digestibility at increasing levels of up to 2000 FTU/kg of liquid phytase. Ingelmann et al. (2019)Ingelmann, C. J.; Witzig, M.; Möhring, J.; Schollenberger, M.; Kühn, I. and Rodehutscord, M. 2019. Phytate degradation and phosphorus digestibility in broilers and turkeys fed different corn sources with or without added phytase. Poultry Science 98:912-922. https://doi.org/10.3382/ps/pey438
https://doi.org/10.3382/ps/pey438...
found that the use of 500 FTU/kg of feed increased pre-caecal phosphorus digestibility by approximately 15% in broilers. Therefore, these data support the results found in the present study.

When supplying 750 FTU/kg of phytase to broiler chickens, Martins et al. (2013)Martins, B. A. B.; Borgatti, L. M. O.; Souza, L. W. O.; Robassini, S. L. D. A. and Albuquerque, R. 2013. Bioavailability and poultry fecal excretion of phosphorus from soybean-based diets supplemented with phytase. Revista Brasileira de Zootecnia 42:174-182. https://doi.org/10.1590/S1516-35982013000300005
https://doi.org/10.1590/S1516-3598201300...
observed a reduction of 12.7% in fecal phosphorus excretion without compromising animal performance. However, other studies reported a reduction of 33% (Applegate et al., 2003Applegate, T. J.; Angel, R. and Classen, H. L. 2003. Effect of dietary calcium, 25- dihydroxycholecalciferol, or bird strain on small intestinal phytase activity in broiler chickens. Poultry Science 82:1140-1148. https://doi.org/10.1093/ps/82.7.1140
https://doi.org/10.1093/ps/82.7.1140...
) and 55% (Laurentiz et al., 2009Laurentiz, A. C.; Junqueira, O. M.; Filardi, R. S.; Duarte, K. F.; Assuena, V. and Sgavioli, S. 2009. Desempenho, composição da cama, das tíbias, do fígado e das excretas de frangos de corte alimentados com rações contendo fitase e baixos níveis de fósforo. Revista Brasileira de Zootecnia 38:1938-1947. https://doi.org/10.1590/S1516-35982009001000012
https://doi.org/10.1590/S1516-3598200900...
) in excreted phosphorus. However, in addition to the use of phytase, the authors also used the strategy of reducing dietary phosphorus. Thus, by combining the two strategies, the improvements in the reduction of excretion were significantly higher. However, in the present study, using only a phosphorus reduction strategy, we found a 35.05% reduction in phosphorus at the 500 FTU dose of phytase and a 44.19% at the 500 FTU + 3000 IU xylanase dose compared with the control. Therefore, xylanase also contributed to a decrease of phosphorus excretion in the environment.

Pontes et al. (2021)Pontes, T. C.; Brito, J. M.; Wernick, B.; Urbich, A. V.; Panaczevicz, P. A. P.; Miranda, J. A. G.; Furuya, V. R. B. and Furuya, W. M. 2021. Top-sprayed phytase enhances the digestibility of energy, protein, amino acids and minerals, and reduces phosphorus output in Nile tilapia fed all-vegetable diets. Aquaculture Research 52:6562-6570. https://doi.org/10.1111/are.15527
https://doi.org/10.1111/are.15527...
provided spray phytase in extruded diets for Nile tilapia, and confirmed that supplementation of this enzyme increases energy and nutrient digestibility, including aminoacids, phosphorus, and cationic minerals. Furthermore, the results were dose-dependent, which corroborates those found in our study. Also, the authors pointed out that phytase is a useful strategy to reduce the inclusion of inorganic phosphorus in diets and nitrogen. In this scenario, phytate increases the mucin section in the intestine and this increases the endogenous loss of aminoacids (Selle et al., 2012Selle, P. H.; Cowieson, A. J.; Cowieson, N. P. and Ravindran, V. 2012. Protein-phytate interactions in pig and poultry nutrition: a reappraisal. Nutrition Research Reviews 25:1-17. https://doi.org/10.1017/S0954422411000151
https://doi.org/10.1017/S095442241100015...
). In addition, phytate binds to cations such as Ca, Mg, Zn, Fe, and Cu, and the addition of phytase can break this complex and provide greater absorption of these minerals (Adeola and Cowieson, 2011Adeola, O. and Cowieson, A. J. 2011. Board-invited review: Opportunities and challenges in using exogenous enzymes to improve nonruminant animal production. Journal of Animal Science 89:3189-3218. https://doi.org/10.2527/jas.2010-3715
https://doi.org/10.2527/jas.2010-3715...
). However, we found no difference in the digestibility of calcium as expected. This could be due to a lower concentration of minerals linked to phytic acid, such as calcium in this case, and also due to the retention time and pH of the gastrointestinal tract that may have influenced the action of the enzyme on the release of minerals. Furthermore, another possible explanation could be the high levels of calcium in the feed, which could have reduced its absorption in the intestine, even with the addition of phytase. Perhaps there was a need to reduce the inclusion of this mineral in the feed so that a difference in its digestibility could be observed.

In addition, most studies that used the two associated enzymes did not demonstrate an improvement in digestibility (Olukosi and Adeola, 2008Olukosi, O. A. and Adeola, O. 2008. Whole body nutrient accretion, growth performance and total tract nutrient retention responses of broilers to supplementation of xylanase and phytase individually or in combination in wheat-soybean meal based diets. The Journal of Poultry Science 45:192-198. https://doi.org/10.2141/jpsa.45.192
https://doi.org/10.2141/jpsa.45.192...
; Olukosi et al., 2020Olukosi, O. A.; González-Ortiz, G.; Whitfield, H. and Bedford, M. R. 2020. Comparative aspects of phytase and xylanase effects on performance, mineral digestibility, and ileal phytate degradation in broilers and turkeys. Poultry Science 99:1528-1539. https://doi.org/10.1016/j.psj.2019.11.018
https://doi.org/10.1016/j.psj.2019.11.01...
), unlike what was seen in the present study, in which there was an improvement in the apparent digestibility coefficient of phosphorus and lower phosphorus excretion when using the association of liquid phytase + xylanase after feed pelletization. Therefore, both the use of isolated liquid phytase and that associated with liquid xylanase after pelletization were efficient in improving the use of phytic phosphorus present in the ingredients of plant origin. This fact may have happened because of the effect of temperature and humidity in the pelletization process, which causes expansion of starch granules (Cruz et al., 2020Cruz, T. A.; Stresser, A. C. P.; Oliveira, S. G.; Almeida, L. M.; Santos, M. C. and Félix, A. P. 2020. Exogenous enzymes and pelleting increase diet digestibility of piglets. Archives of Veterinary Science 25:87-94. https://doi.org/10.5380/avs.v25i1.68274
https://doi.org/10.5380/avs.v25i1.68274...
), and thus facilitates access of the enzymes to substrates, such as NSP and phytate.

5. Conclusions

The addition of phytase alone or in combination with xylanase is able to improve the apparent digestibility and reduce phosphorus excretion. Therefore, the use of liquid phytase and liquid phytase + xylanase after pelletization becomes interesting to reduce the inclusion of inorganic phosphorus, which can reduce the feed cost and phosphorus excretion in the environment. Furthermore, applying the enzymes after pelleting is a useful alternative since it prevents denaturation or inactivation of the enzymes.

References

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Publication Dates

  • Publication in this collection
    09 Oct 2023
  • Date of issue
    2023

History

  • Received
    19 Apr 2022
  • Accepted
    09 Feb 2023
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