Effect of a protease on the ileal and fecal digestibility of amino acids of canola meal and dry distillers grains with solubles in starter broilers
Published: January 30, 2012
By: S Fernández Tinoco (DSM Nutritional Products México SA de CV), S Gómez Rosales, ML Angeles, E Ramírez (CENIDFyMA-INIFAP) (FESC-UNAM)
Summary
An experiment was carried out to evaluate the effect of a protease on the ileal and fecal apparent digestibility (IAD and FAD) of amino acids (AA) and the apparent metabolizable energy corrected to cero nitrogen retention (AMEn) in canola meal (CM) and dry distillers grain with solubles (DDGS) in broilers from 13 to 21 d of age. Three hundred and sixty chicks were assigned to 6 treatments in a complete randomized design with a factorial arrangement of 3 dietary treatments x 2 levels of a protease (Ronozyme ProAct: 0 and 200 g/ton). The dietary treatments were as follows: 1. Basal diet: formulated with corn and soybean meal and nutrient content according to the recommendations of Ross for broilers from 15 to 28 days of age (Basal); 2. As 1) plus protease 3. 30% of the basal diet was replaced by 30% CM; 4. As 3) plus protease 5. 30% of the basal diet was replaced by 30% DDGS; and 6. As 5) plus protease. The IAD and FAD of AA were estimated using the substitution method. Results were subjected to ANOVA. The IAD and FAD of some AA were affected by the Diet and CM and DDGS. The average FAD of Lysine across all diets as well as the FAD of Lysine in CM was improved by Protease. The IAD and FAD of the rest AA was also arithmetically improved by Protease. In addition, Protease improved the AMEn of all diets and CAM and DDGS. In summary, the results suggest that the availability of AA and the energetic value of diets formulated with CM and DDGS could be enhanced by the addition of a protease.
Key Words: Starter broilers, Protease, Canola meal, DDGS, Amino acids.
Introduction
Individual feed ingredients commonly used in poultry may vary widely in composition and nutrient availability because of variation in cultivars, growing conditions, processing and storage conditions (NRC, 1994). In spite of this, in recent years the major improvements in nutrition and the efficiency of production have relied on obtaining maximum nutrient utilization from feedstuffs, mainly by using exogenous enzymes directed to different chemical fractions such as starch and non-starch polysaccharide (NSP) (Slominski, 2010). By using exogenous enzymes, improvements on nutrient digestibility such as phosphorus (P), calcium (Ca), fat, starch and NSP have been observed as well as for the energy (E) value of diets and feedstuffs. Another advantage of the addition of exogenous enzymes is the improvement of the nutritional value of some ingredients currently considerer of lower quality such as canola meal (CM) and dry distillers grains with solubles (DDGS) when compared to soybean meal (SBM). CM and DDGS have lower content and digestibility of AA and higher content of NSP compared to SBM, which have limited their broad use in poultry feeding. In CM and DDGS, the availability of P, Ca and E have been improved by dietary supplementation of phytase, glucanase and xylanase in poultry and swine diets (Gómez et al., 2009a; 2009b; Angeles & Gómez, 2010; Gómez & Angeles, 2010). However, it remains to clarify whether the addition of a protease enzyme can improve the feeding value of CM and DDGS by increasing the digestibility of AA. Therefore, an experiment was carried out to evaluate the effect of a protease on the apparent ileal and fecal AA digestibility of CM and DDGS in broilers from 13 to 21 d of age.
Materials & Methods
Three hundred and sixty 9 day old, male B308 Ross chicks, were allocated in groups of three in individual crates provided with a metal feeder and a cup drinker. From day 9 to 12, chicks were allowed to adapt to the crates and feeding schedule. Chicks were fed ad libitum, but feed was served three times during the day to minimize feed wastage. The diet offered during this period was formulated according to the nutrient recommendations of Ross 308 Management Guide for broilers from 1 to 14 days of age. At day 13, chicks were randomly assigned to 8 treatments in a complete randomized design with a factorial arrangement of 3 dietary treatments x 2 levels of a protease (Ronozyme ProAct). The dietary treatments were as follows:
1. Basal diet: formulated with corn and soybean meal and nutrient content according to the recommendations of Ross for broilers from 15 to 28 days of age. The dietary nutrient content was as follows: 3141 kcal of ME/kg of feed, 1.05% of digestible lysine, 0.90% of Ca and 0.45% of Available P (Basal).
2. As 1) plus 200 g/ton of protease
3. 30% of the basal diet was replaced by 30% CM
4. As 3) plus 200 g/ton of protease
5. 30% of the basal diet was replaced by 30% DDGS
6. As 5) plus 200 g/ton of protease
2. As 1) plus 200 g/ton of protease
3. 30% of the basal diet was replaced by 30% CM
4. As 3) plus 200 g/ton of protease
5. 30% of the basal diet was replaced by 30% DDGS
6. As 5) plus 200 g/ton of protease
Broilers were adapted to the experimental diets during 4 days. Feed was added with 0.25% of titanium oxide and was offered on an ad libitum basis, in three meals per day. Feces were totally collected, every 12 h, during the last four days of the test period, and stored frozen. The last day of the experiment, all chicks were killed, by cervical dislocation, the ileon was exteriorized, from the Meckel diverticulum to the ileocecal valve; then the ileal content was gently rinsed with distilled water into plastic bags and immediately frozen.
Laboratory analysis: the ileal content and feces were unfrozen, lyophilized, ground and kept at 4o C. In feed samples and ileal content dry matter (DM), nitrogen (N), AA and titanium were determined following the recommendations of the AOAC (1994). Diets and total fecal samples collected for AA determinations were also processed in the lab and analyzed for DM, N and E. Then, the nutrient retention was estimated. In addition, the AMEn was calculated for the diets and feed ingredients. To estimate the ileal or fecal output of AA and the IAD and FAD of AA the recommendation of Kadim et al. (2002) were followed.
Results were subjected to analysis of varianza using the GLM procedures of SAS. To analyze the diets, six treatments were considered and to analyze the feed ingredients, four treatments were considered in the models. Then, each feed ingredient was analyzed individually without and with Protease. The ileal content and fecal samples of two replicate crates per treatment were pooled to get one sample for lab analysis. Therefore, the experimental unit was two adjacent crates of the same treatment with a total of six chicks. There were five replicates per treatment.
Results & Discussion
The IAD of AA
The interaction between Diet or Feed ingredients and Protease did not show any statistical difference on the IAD of AA. The Diets showed differences on the IAD on some AA: The IAD of Lysine was higher (P < 0.01) on the Basal and Basal + CM and lower on the Basal + DDGS diet. The IAD of Tryptophane and Threonine were higher (P < 0.01) on the Basal than for the Basal + CM and Basal + DDGS diets. The IAD of Arginine was higher (P < 0.01) on the Basal diet, intermediate on the Basal + CM and lower on the Basal + DDGS diet. The Feed ingredients also had a significant effect on some AA: The IAD of Lysine and Arginine were higher (P < 0.01) on CM and lower on DDGS. There was an arithmetic improvement on the IAD with Protease addition on the Diets and Feed ingredients for all AA, but none of these reached statistical differences.
The interaction between Diet or Feed ingredients and Protease did not show any statistical difference on the IAD of AA. The Diets showed differences on the IAD on some AA: The IAD of Lysine was higher (P < 0.01) on the Basal and Basal + CM and lower on the Basal + DDGS diet. The IAD of Tryptophane and Threonine were higher (P < 0.01) on the Basal than for the Basal + CM and Basal + DDGS diets. The IAD of Arginine was higher (P < 0.01) on the Basal diet, intermediate on the Basal + CM and lower on the Basal + DDGS diet. The Feed ingredients also had a significant effect on some AA: The IAD of Lysine and Arginine were higher (P < 0.01) on CM and lower on DDGS. There was an arithmetic improvement on the IAD with Protease addition on the Diets and Feed ingredients for all AA, but none of these reached statistical differences.
The FAD of AA
The Diets showed differences on the IAD on some AA: The FAD of Lysine and Methionine were higher (P < 0.01) on the Basal + CM diet than the Basal + DDGS diet. The FAD of Arginine was higher (P < 0.05) on the Basal + CM compared to the Basal + DDGS diet. The FAD of Leucine was higher (P < 0.05) on the Basal diet compared to the Basal + CM and Basal + DDGS diets. The FAD of Phenylalanine was higher (P < 0.01) on the Basal and Basal + DDGS diets compared to the Basal + CM. The Feed ingredients also had a significant effect on some AA: The FAD of Lysine, Methionine and Arginine were higher (P < 0.01) on CM compared to DDGS. The Phenylalanine was higher for DDGS than for CM (P < 0.05). The average FAD of Lysine (over all diets) showed a trend for being higher (P < 0.10) with the addition of Protease. The addition of Protease to the Feed ingredients caused an increase on the average FAD of Lysine (P < 0.05) and Arginine (P < 0.10) and also increased the FAD of Lysine on CM. For the rest AA there was an arithmetic improvement on the FAD with Protease addition on the Diets and Feed ingredients.
The total, ileal and fecal apparent digestible AA concentration on CM and DDGS were, in most cases, inside the range of values reported in the literature. On the diets, the addition of Protease improved the IAD of Methionine, Cystine, Threonine, Phenylalanine and Tyrosine by 1-2%; and Lysine, Isoleucine, Valine, Leucine and Histidine by 0.7-0.9%. On the feed ingredients, the addition of Protease improved the IAD of Lysine, Methionine and Cystine by 3-4%; Tryptophane, Threonine, Valine, Histidine, Phenylalanine and Tyrosine by 2-3%; and, Arginine and Isoleucine by 0.5-1%. On the diets, the addition of Protease improved the fecal digestibility of Lysine, Cystine, Arginine, Isoleucine, Valine, Histidine and Phenylalanine by 1-2%; and Methionine, Tryptophane, Threonine, Leucine and Tyrosine by 0.6-0.95%. On the feed ingredients, the addition of Protease improved the fecal digestibility of Lysine, Isoleucine and Phenylalanine by 3-5%; Methionine, Cystine, Arginine, Valine, Histidine and Tyrosine by 2-3%, and; Tryptophane, Threonine and Leucine by 1-2%.
In addition, Protease improved the AMEn on the diets by 1% and on the feed ingredients by 3.3%.
It could be noted that in some AA the ileal digestibility was improved by 3-4% and the fecal digestibility was improved by 4-5% on CM and DDGS. These values are close to some of the results presented previously with the use of the same Protease in broiler chickens. The lack of statistical differences on most of the AA evaluated on the present study could be due to the low number of replicates (5) compared to previous studies with Protease in which 7-9 replications were used per treatment (Maiorka et al., 2009; Vieira et al., 2009; Angel et al., 2010). Other differences seen with previous studies were the type of diet used (corn-soybean meal-meat bone meal) and that the AA digestibilities were determined for whole diets. Also, in the present study, feed was restricted whereas on the others, broilers were fed ad libitum. On the experiments of Maiorka et al. (2009) and Vieira et al. (2009) ileal contents were collected on birds of 42 and 40 d of age, respectively. On the study of Angel et al. (2010) chicks were fed from 7 to 22 d of age and ileal content was removed at 22 d of age, similar to the procedures of the present trial, however, Protease was added to a negative control low-CP diet added with crystalline amino acids.
The Diets showed differences on the IAD on some AA: The FAD of Lysine and Methionine were higher (P < 0.01) on the Basal + CM diet than the Basal + DDGS diet. The FAD of Arginine was higher (P < 0.05) on the Basal + CM compared to the Basal + DDGS diet. The FAD of Leucine was higher (P < 0.05) on the Basal diet compared to the Basal + CM and Basal + DDGS diets. The FAD of Phenylalanine was higher (P < 0.01) on the Basal and Basal + DDGS diets compared to the Basal + CM. The Feed ingredients also had a significant effect on some AA: The FAD of Lysine, Methionine and Arginine were higher (P < 0.01) on CM compared to DDGS. The Phenylalanine was higher for DDGS than for CM (P < 0.05). The average FAD of Lysine (over all diets) showed a trend for being higher (P < 0.10) with the addition of Protease. The addition of Protease to the Feed ingredients caused an increase on the average FAD of Lysine (P < 0.05) and Arginine (P < 0.10) and also increased the FAD of Lysine on CM. For the rest AA there was an arithmetic improvement on the FAD with Protease addition on the Diets and Feed ingredients.
The total, ileal and fecal apparent digestible AA concentration on CM and DDGS were, in most cases, inside the range of values reported in the literature. On the diets, the addition of Protease improved the IAD of Methionine, Cystine, Threonine, Phenylalanine and Tyrosine by 1-2%; and Lysine, Isoleucine, Valine, Leucine and Histidine by 0.7-0.9%. On the feed ingredients, the addition of Protease improved the IAD of Lysine, Methionine and Cystine by 3-4%; Tryptophane, Threonine, Valine, Histidine, Phenylalanine and Tyrosine by 2-3%; and, Arginine and Isoleucine by 0.5-1%. On the diets, the addition of Protease improved the fecal digestibility of Lysine, Cystine, Arginine, Isoleucine, Valine, Histidine and Phenylalanine by 1-2%; and Methionine, Tryptophane, Threonine, Leucine and Tyrosine by 0.6-0.95%. On the feed ingredients, the addition of Protease improved the fecal digestibility of Lysine, Isoleucine and Phenylalanine by 3-5%; Methionine, Cystine, Arginine, Valine, Histidine and Tyrosine by 2-3%, and; Tryptophane, Threonine and Leucine by 1-2%.
In addition, Protease improved the AMEn on the diets by 1% and on the feed ingredients by 3.3%.
It could be noted that in some AA the ileal digestibility was improved by 3-4% and the fecal digestibility was improved by 4-5% on CM and DDGS. These values are close to some of the results presented previously with the use of the same Protease in broiler chickens. The lack of statistical differences on most of the AA evaluated on the present study could be due to the low number of replicates (5) compared to previous studies with Protease in which 7-9 replications were used per treatment (Maiorka et al., 2009; Vieira et al., 2009; Angel et al., 2010). Other differences seen with previous studies were the type of diet used (corn-soybean meal-meat bone meal) and that the AA digestibilities were determined for whole diets. Also, in the present study, feed was restricted whereas on the others, broilers were fed ad libitum. On the experiments of Maiorka et al. (2009) and Vieira et al. (2009) ileal contents were collected on birds of 42 and 40 d of age, respectively. On the study of Angel et al. (2010) chicks were fed from 7 to 22 d of age and ileal content was removed at 22 d of age, similar to the procedures of the present trial, however, Protease was added to a negative control low-CP diet added with crystalline amino acids.
Nutrient retention and AMEn
The DM excretion was lower (P < 0.01) and the DM retention was higher (P < 0.01) on the Basal diet related to the rest of the diets. The N excretion was lower on the Basal and higher on the Basal + CM and Basal + DDGS diets (P < 0.01). The N retention was higher on the Basal, intermediate on the Basal + CM and lower on the Basal + DDGS (P < 0.01). The energy excretion was lower (P < 0.01) on the Basal compared to CM and DDGS diets. The AMEn was not statistically different among diets (3037, 2775 and 2976 kcal/kg, SEM= 24.105). Protease did not affect any of the response variables evaluated, however, the addition of Protease arithmetically improved the energy released by all diets as follows: 60, 32 and 27 kcal/kg of the Basal, Basal + CM and Basal + DDGS, respectively.
The AMEn differed among Feed ingredients. The AMEn was lower in CM than for DDGS (2234 and 2904 kcal/kg, SEM= 43.40; P < 0.01). The addition of Protease did not statistically affect the AMEn on the ingredients; however, it did numerically improve the energy released as follows: 106 and 89 kcal/kg for CM and DDGS, respectively.
The DM excretion was lower (P < 0.01) and the DM retention was higher (P < 0.01) on the Basal diet related to the rest of the diets. The N excretion was lower on the Basal and higher on the Basal + CM and Basal + DDGS diets (P < 0.01). The N retention was higher on the Basal, intermediate on the Basal + CM and lower on the Basal + DDGS (P < 0.01). The energy excretion was lower (P < 0.01) on the Basal compared to CM and DDGS diets. The AMEn was not statistically different among diets (3037, 2775 and 2976 kcal/kg, SEM= 24.105). Protease did not affect any of the response variables evaluated, however, the addition of Protease arithmetically improved the energy released by all diets as follows: 60, 32 and 27 kcal/kg of the Basal, Basal + CM and Basal + DDGS, respectively.
The AMEn differed among Feed ingredients. The AMEn was lower in CM than for DDGS (2234 and 2904 kcal/kg, SEM= 43.40; P < 0.01). The addition of Protease did not statistically affect the AMEn on the ingredients; however, it did numerically improve the energy released as follows: 106 and 89 kcal/kg for CM and DDGS, respectively.
Conclusions
The ileal apparent digestibility of AA on the Diets and on CM and DDGS were not affected statistically by the addition of Protease, but in all cases, there were arithmetic improvements with the addition of the enzyme. The average fecal apparent digestibility of Lysine was improved across all diets by Protease as well as the fecal apparent digestibility of Lysine in CM. The digestibility of the rest AA was also arithmetically improved by Protease. In addition, Protease improved the AMEn of all the diets and Feed ingredients.
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