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Article

Enzyme Supplementation to Diets Containing Wheat Distillers’ Dried Grains with Solubles (DDGS) When Fed to Laying Hens

by
Isobel M. Whiting
1,*,
Stephen P. Rose
1,
Ahmed M. Amerah
2 and
Vasil R. Pirgozliev
1
1
The National Institute of Poultry Husbandry, Harper Adams University, Shropshire TF10 8NB, UK
2
Danisco Animal Nutrition, Marlborough, Wiltshire SN8 1XN, UK
*
Author to whom correspondence should be addressed.
Poultry 2025, 4(2), 22; https://doi.org/10.3390/poultry4020022
Submission received: 18 March 2025 / Revised: 2 April 2025 / Accepted: 24 April 2025 / Published: 6 May 2025
Versions Notes

Abstract

:
A study was conducted using 144 Hy-Line Brown laying hens (22 weeks old) to assess the impact of exogenous enzymes on energy utilisation and ileal nutrient digestibility in diets containing 300 g/kg wheat distillers’ dried grains with solubles (DDGS). A basal diet was prepared and divided into eight treatments: a control (C) and diets supplemented with 2000 units/kg xylanase (XYL), 500 units/kg phytase (PHY), and 4000 units/kg protease (PRO), individually and in combination. The diets were fed for nine days to six coops, each housing three birds. Feed intake, weight gain, and feed conversion ratio were recorded. The feed and excreta were analysed for gross energy, and the apparent metabolisable energy (AME) was calculated. On the final day, the birds were euthanised, and ileal digesta were collected, freeze-dried, and analysed for the digestibility coefficients of dry matter (DM), nitrogen, fat, and neutral detergent fibres (NDFs). XYL supplementation improved (p = 0.035) dietary AME but did not affect (p > 0.05) DM, nitrogen, fat, or NDF digestibility. No significant effects (p > 0.05) were observed for PHY or PRO, and no interactions (p > 0.05) were found between enzyme combinations. Substrates in experimental diets involving various enzyme combinations should be given careful consideration.

1. Introduction

Distillers’ dried grains with solubles (DDGS) are a byproduct of bioethanol production. In the UK, wheat is the predominant choice of cereal grain used. The subsequent conversion of starch to ethanol results in the protein, phytate, and fibre content of the DDGS becoming concentrated by approximately threefold [1,2]. In addition to the increased nutrient profile, DDGS have been found to contain a high content of non-starch polysaccharides (NSPs) [3,4]. Poultry are not able to digest NSPs, as birds lack the required endogenous enzymes. As a result, the absorptive and digestive dynamics of the gut affect the overall nutrient uptake [5]. Over the past few decades, exogenous enzymes have been used in poultry diets to obtain productive uniformity by improving energy utilisation and nutrient digestibility [6]. Information on the use of supplemental enzymes in diets containing wheat DDGS will be an important contribution for poultry nutritionists. Previous research has advised using NSP-degrading enzymes to alleviate the negative effects imposed by highly fibrous feeds [7,8,9], such as DDGS. The simultaneous use of exogenous enzymes such as carbohydrases, e.g., xylanases (XYLs), phytases (PHYs), and proteases (PROs), has been found to have not only additive and synergistic, but also antagonistic effects when added to broiler diets [8,10]. Phytases are routinely used in poultry feeds to improve P availability; in addition, they have also been found to improve the availability of both energy and amino acids [11]. Despite birds being able to digest the majority of dietary protein, as much as 10% can escape digestion in the small intestine [8]. Protein digestibility has been observed to improve when exogenous PROs are supplemented into diets [12,13]. Although there is a reason to expect an improved production performance when using a combination of exogenous enzymes [8], this is not always the case. Enzymes work on substrates; thus, the efficacy of each enzyme would depend on the presence of a specific substrate in the diet [6]. The dynamics of substrate hydrolysis in a diet may also bring unclarity in the modes of action of exogenous enzymes [1,7]. In addition, there are many interactions between the supplementary enzymes and the host, e.g., intestinal microflora, which require further investigation [8,10]. The poultry industry needs more certainty when using a mixture of different exogenous enzymes in diets. Thus, there is a need to generate more practical information on various enzyme combinations and dietary formulations, allowing for a better understanding of the mode of action of enzymes. A diet containing relatively high levels of fibre, phytate, and poorly digestible protein may be a good model to test the interaction between the enzymes. The aim of the present experiment was to feed laying hens diets containing 300 g/kg wheat DDGS with the addition of exogenous XYL, PHY, and PRO, both singularly and in combination.

2. Materials and Methods

2.1. Diet Formulation

A basal feed (Table 1) with 495.6 g/kg wheat, 300 g/kg wheat DDGS, and 50.0 g/kg soybean meal as the main ingredients, contained 11.40 MJ/kg apparent metabolizable energy (AME) and 171.1 g/kg crude protein. The basal feed was split into 8 parts, and 1 part was fed as is, as an unsupplemented control, and the remaining seven parts were supplemented with exogenous XYL (Danisco Xylanase), PHY (Phyzyme® XP), and PRO (Axtra PRO), both singularly and in combination, resulting in a total of 8 diets.
All enzymes were developed by Danisco Animal Nutrition (DuPont Industrial Biosciences, Marlborough, UK). Danisco XYL (EC 3.2.1.8) is a preparation of endo-1,4-ß-XYL produced from a species of fungus called Trichoderma reesei. Phyzyme® XP (EC 3.1.3.26) is a bacterial 6-PHY, sourced from a species of Escherichia coli, and is expressed in Saccharomyces pombe. Axtra PRO (EC 3.4.21.62) is a subtilisin PRO, expressed in Bacillus subtilis. The enzymes were added on top of the feed, in powder form, at 0.250 g/kg, providing activity of 2000, 500, and 4000 (U/kg) for XYL, PHY, and PRO, respectively. The diets did not contain any coccidiostat, antimicrobial growth promoters, prophylactic, or other similar additives. All of the diets were fed as a mash with 5 g/kg titanium dioxide (TiO2) added on top as an indigestible marker. From sixteen to twenty-two weeks of age, the birds were fed a basal feed only, without any enzymes, coccidiostat, antimicrobial growth promoters, prophylactic, or other similar additives. Then, the experimental diets were administered for nine days with six replicates per treatment.

2.2. Bird Husbandry

The study, project number 10-201502, was approved by the Animal Experimental Committee of Harper Adams University. A total of 144 sixteen-week-old Hy-Line Brown laying hens were obtained from a commercial supplier (Country Fresh Pullets Ltd., Oswestry, Shropshire, UK). The birds were randomly allocated to 48 layer coops (over three tiers) in groups of three. Each coop was equipped with a separate feeder at the front and two nipple drinkers inside. Coop dimensions were 45 cm × 40 cm × 50 cm and consisted of wire mesh flooring, which contained no bedding material. Temperature was maintained at 21 °C and relative humidity was between 50 and 70%. The birds had ad libitum access to feed and water. The lighting was set to a 10 h day length upon arrival. At 20 weeks of age, the lighting was increased by 30 min each week until it reached 16 h per 24 h period. The nine-day feeding experiment started when the birds were 22 weeks old (body weight 1665.1 g STDEV ± 77.39). Feed intake was measured for the entire experimental period. Excreta were collected every day for the last four experimental days, oven-dried at 60 °C, milled to pass through a 0.5 mm mesh, and their chemical composition and gross energy (GE) were determined to calculate dietary apparent metabolisable energy (AME). On the final day of the study, the birds were killed by cervical dislocation, and digesta were collected from the ileum, between Meckel’s diverticulum and the caecal junction, and pooled into separate pots per cage. The ileal digestibility coefficients of dietary dry matter (DM), nitrogen, neutral detergent fibres (NDFs), and fat were then determined.
Digesta were immediately frozen at −20 °C for 24 h and then freeze-dried and milled using a 0.5 mm mesh. Using the marker technique, the digesta were used to determine nutrient digestibility. In addition, some hen performance variables, including feed intake (FI), body weight gain (WG), egg production (eggs per hen day), egg mass, and feed conversion ratio (FCR) for egg production, were recorded as control measures of baseline performance metrics.

2.3. Chemical Analyses

The dry matter (DM), ash, crude protein (CP) (N × 6.25), and oil (as ether extract) in the samples were determined following a standard methodology [14,15,16,17]. The gross energy in the feed and excreta was determined using a bomb calorimeter (Parr 6200 Instrument Company, Moline, IL, USA, 61,265) as described elsewhere [18,19]. The neutral detergent fibre (NDF) [20], non-starch polysaccharides (NSPs) [21], and TiO2 [22] in the samples were also determined. The enzyme activities in the feed samples were measured at the DuPont Nutrition Biosciences Innovation Laboratories (Brabrand, Denmark). One phytase unit was defined as the quantity of enzyme that releases 1 µmol of inorganic P/min from 5.0 mM sodium phytate at pH 5.5 at 37 °C. One xylanase unit was defined as the amount of enzyme that releases 0.48 µmol of reducing sugar as xylose from wheat arabino xylan per minute at pH 4.2 and 50 °C. One protease unit was defined as the amount of enzyme that releases 1.0 µg of phenolic compound, expressed as tyrosine equivalents, from a casein substrate per minute at pH 7.5 and 40 °C. All samples were analysed in technical duplicates.

2.4. Calculations

The AME of the diets was determined by the total collection technique [23], measuring the GE of the diet consumed and the amount of GE excreted and calculating the amount per kilogram of feed.
Dietary AME (MJ/kg) was calculated as:
A M E = G E   i n t − G E   o u t F e e d   i n t a k e
where AME (MJ/kg) = the apparent metabolizable energy content of the diet, and GE = the gross energy of the diet and excreta, respectively; GE int = the GE of the diet multiplied by the feed intake; GE out = the GE of the excreta multiplied by the excreta voided; Feed intake = the feed intake during the collection period (kg).
The ileal nutrient retention coefficients were calculated using the following equation:
N u t r i e n t   r e t e n t i o n = N / T i D i e t   − N / T i D i g e s t a ( N / T i ) D i e t  
where (N/Ti) Diet = the ratios of DM, nitrogen, fat, or NDF to the TiO2 in the diet, and (N/Ti) Digesta = the ratios of DM, nitrogen, fat, or NDF to the TiO2 in the digesta samples [24].
F C R   e g g   p r o d u c t i o n = F e e d   i n t a k e E g g   m a s s
The feed conversion ratio (FCR) for egg production was calculated by dividing the feed intake per hen over the study period by the egg mass for the same period.

2.5. Statistical Analysis

Statistical comparisons were performed using the general ANOVA procedure of Genstat 23rd edition (VSN International Ltd., Hemel Hempstead, UK) in a 2 × 2 × 2 factorial arrangement testing for the main effects of XYL, PHY, PRO, and the interaction term. Additionally, the unsupplemented control and the rest of the diets were compared with a single contrast comparison test within the factorial ANOVA model. All data were checked for the normality and homogeneity of the residuals prior to ANOVA.

3. Results and Discussion

The objective of this experiment was to evaluate whether the nutritional value of a diet containing a high level of wheat DDGS could be improved by the addition of exogenous enzymes. The determined greater CP and fat dietary contents (Table 1) may be explained by the differences in the CP composition in the used raw materials and the information in the dietary formulation software. Table 2 shows the proximate analysis of the studied wheat DDGS. The results from the proximate analysis are generally in agreement with data published by others [1,2,25,26].
The recovery of the supplementary dietary enzymes is presented in Table 3. There was a range of enzyme activities within the diets, as the XYL and PHY × PRO diets contained about 56% more XYL and PHY enzymes than expected, respectively.
The effects of the different treatments on the AME and nutrient digestibility are presented in Table 4. The addition of exogenous XYL improved (p = 0.035) AME by 0.59 MJ/kg. No effects (p > 0.05) were observed for PHY and PRO or the enzyme interactions for any of the other variables measured.
The results in Table 5 show that there were no abnormalities regarding hen performance variables during the study. The mean daily weight gain of 3 g per bird was within the expected range for those ages of Hy-Line Brown hens [27]. Romero et al. [28] reported slightly greater egg mass and lower FCR possibly due to the use of elderly Hy Line layers and employing a longer study period.

3.1. The Effect of Xylanase on the Studied Wheat DDGS

Most research on the impact of supplementary enzymes on the feeding quality of bioethanol co-products has focused on maize DDGS [29,30]. Since wheat contains higher levels of NSP than maize, it is reasonable to expect a greater improvement in the feeding value of wheat DDGS when diets are supplemented with exogenous enzymes such as xylanase. [31]. The addition of XYL to broiler diets has been found to hydrolyze the high molecular mass sub-fraction of arabinoxylan and reduce the formation of viscous solutions in the digestive tract, thus improving energy and nutrient availability [31]. There is a negative relationship between dietary AME and soluble NSP, as this NSP fraction has a high water-holding capacity and increases digesta viscosity [31,32]. The increase in dietary AME with XYL supplementation may indicate that the enzyme hydrolyzed soluble NSP in the gut, thereby preventing the formation of a highly viscous environment in the digesta. Similar studies have also reported improvements in energy utilisation in wheat-based DDGS diets containing exogenous XYL for laying hens [32] and broilers [33,34,35]. The improved dietary AME may also be due to potentially more available substrate for XYL enzyme in this study.

3.2. The Effect of PHY on the Studied Wheat DDGS

It was indicated by Martinez-Amezcua et al. [36] that the bioavailability of phosphorus in maize DDGS for broiler chicks could be considerably improved with the addition of microbial PHY. Phytase acts by dephosphorylating phytate, enabling the release of phosphorus, as well as other nutrients that phytate may have complexed with [37,38,39]. In this experiment, the addition of PHY had no effect on any of the parameters measured. The lack of an effect of PHY may be due to the fact that the diets were formulated to contain a sufficient level of phosphorus. In addition, observations made by Liu et al. [40] indicated that phytate is, to some extent, degraded during fermentation by the action of yeast PHY, thus increasing the bioavailability of phosphorus in DDGS, consequently potentially reducing the efficacy of the exogenous PHY. Furthermore, the efficacy of PHY has been found to vary depending on grain type. When investigating the effects of exogenous PHY, Liu et al. [41] observed more robust responses, particularly for nutrient utilisation, in maize-based feeds compared with wheat and sorghum. Similar findings were also reported by Truong et al. [11].

3.3. The Effect of PRO on the Studied Wheat DDGS

Hens are unable to produce enough endogenous XYL and PHY in their gastrointestinal tract, but they can synthesise sufficient amounts of PRO to help feed protein utilisation [42]. However, despite birds being able to digest the majority of dietary protein, a significant amount, e.g., about 10% can escape digestion in the small intestine [8,43]. Exogenous PRO can be used in poultry diets to improve protein digestibility [13]. Indeed, apparent ileal digestibility has been observed to improve when exogenous PRO is supplemented into diets [13,44]. Improvements in the nutrient digestibility and performance of broilers have also been reported following the supplementation of exogenous PRO alone and in combination with other exogenous enzymes, such as PHY and XYL [45,46]. However, the lack of an effect of exogenous PRO among energy and nutrients in the present study may be because the birds produced a sufficient level of endogenous PRO. In addition, the DDGS may be heat damaged as a consequence of Maillard reactions occurring during the production process. The Maillard reaction is a form of nonenzymatic browning caused under high temperatures by a chemical reaction occurring between reducing sugars and amino acids. The Maillard reaction is regarded as being primarily responsible for causing chemical heat damage to proteins within DDGS [47,48]. Therefore, if the DDGS used in the present study was subject to Maillard reactions then the protein will be irreversibly bound, rendering it nutritionally unavailable, thus also partly explaining the lack of effect of PRO [49]. In addition, the relatively high CP dietary content may also prevent PRO from having a visible effect on studied variables [13,44,45].

Enzyme Combinations

There is inconsistency in the response of poultry growth performance and nutrient digestibility to exogenous enzyme combinations. Depending on the specific enzyme combination used, responses range from antagonistic [50] to subadditive [51], additive [45,46], and even synergistic [52]. This variability may be attributed to differences in dietary composition, experimental design, bird genotype, and the use of enzyme complexes rather than single-component enzyme preparations [8,11,13]. When several enzymes are tested together the effect of the enzyme preparation cannot always be attributed to the addition of a specific enzyme.
In the present study, the use of the different exogenous enzymes in combination with one another had no effect on any of the variables measured. A study by Waititu et al. [53] looked at the combined addition of XYL, PHY, and PRO to broiler diets containing 5% wheat–maize DDGS. Despite improving feed efficiency and decreasing digesta viscosity, no effect was observed for nutrient digestibility. Similar findings were reported by Barekatain et al. [54], who reported the simultaneous use of XYL and PRO in broiler diets containing 30% sorghum DDGS improved FCR and reduced the concentration of insoluble NSP yet had no effect on nutrient digestibility. Studies undertaken by Min et al. [55,56] found multi-enzyme supplementation to broiler diets containing 30% maize DDGS also had no significant effect on nutrient digestibility, even when added to diets at 1, 2, 3, and 4 times the supplier’s recommended level.
Luo et al. [57] reported that phytate limits nutrient release in field beans and PHY alone or in combination with XYL additively increases their relative nutrient release in an in vitro study. Dietary XYL increases the access of proteolytic enzymes to substrates, e.g., phytate and protein, in the aleurone layer of wheat [58], suggesting that XYL may increase the access of the enzymes to substrates by disrupting the cell wall matrix. Phytic acid and dietary fibre are both responsible for the decreased nutrient availability in diets [59,60]. Thus, PHY might also assist the hydrolytic functions of XYL by dephosphorylation of phytate. The ability of exogenous PRO to increase the solubilisation and fermentation of hemicellulose by removing structural proteins in the cell wall, allowing for the faster access of microbes to digestible substrates has been demonstrated in ruminant models [61]. Again, we did not observe these enzyme interactions in the current study, possibly because diets were providing most of the nutrients needed and there was no room for improvement when enzymes were added, thus suggesting that considering substrates in diets is an important point when incorporating exogenous enzymes.

4. Conclusions

Compared with maize DDGS, very little information is available on the use of exogenous enzymes in poultry diets containing wheat DDGS. However, the addition of non-starch polysaccharide hydrolysing enzymes, such as xylanase to poultry diets containing DDGS, appears to be more efficient compared with phytase and protease. The findings from the present study conclude that the improvement of dietary energy utilisation due to exogenous xylanase has occurred in laying hens. Studies involving various enzyme activities considering substrates in diets are warranted.

Author Contributions

Conceptualization, V.R.P. and S.P.R.; methodology, I.M.W., V.R.P. and S.P.R.; software, I.M.W., V.R.P. and S.P.R.; validation, V.R.P., S.P.R. and A.M.A.; formal analysis, I.M.W.; investigation, I.M.W.; resources, V.R.P.; data curation, I.M.W., V.R.P. and S.P.R. writing—original draft preparation, I.M.W.; writing—review and editing, I.M.W., V.R.P. and S.P.R. visualisation, V.R.P.; supervision, V.R.P.; project administration, I.M.W., V.R.P. and A.M.A.; funding acquisition, V.R.P. and A.M.A. All authors have read and agreed to the published version of the manuscript.

Funding

This research received no external funding.

Institutional Review Board Statement

The animal study protocol was approved by the Ethics Committee of Harper Adams University, project number 10-201502, on 27 March 2015.

Informed Consent Statement

Not applicable.

Data Availability Statement

The original contributions presented in this study are included in the article. Further inquiries can be directed to the corresponding author.

Acknowledgments

The authors would like to acknowledge the technical support provided by Ros Crocker and Richard James.

Conflicts of Interest

The authors declare no conflicts of interest.

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Table 1. The chemical composition of the experimental diets.
Table 1. The chemical composition of the experimental diets.
IngredientAmount g/kg
Wheat495.6
Wheat DDGS300.0
Soya bean meal (48 Crude Protein)50.0
Vegetable oil34.6
Dicalcium phosphate10.0
Limestone102.0
NaCl1.49
Lysine3.8
Methionine2.2
Tryptophane0.3
Vitamin and trace mineral premix 10.01
Calculated composition (as fed basis)
Metabolisable energy (MJ/kg)11.40
Crude protein (g/kg)171.1
Ca (g/kg)42.3
Available P (g/kg)4.5
Analysed values (DM basis) 2
Dry matter (g/kg)882.9
Gross energy (MJ/kg DM)17.85
Crude protein (g/kg DM)192.5
Fat (g/kg DM)64.4
Neutral detergent fibre (g/kg DM)197.4
1 Vitamin and mineral premix provided (units per kg/feed): retinol, 2160 µg; cholecalciferol, 75 µg; α-tocopherol, 25 mg; menadione, 1.5 mg; riboflavin, 5 mg; pantothenic acid, 8 mg; cyanocobalamin, 0.01 mg; pyridoxine, 1.5 mg; thiamine, 1.5 mg; folic acid, 0.5 mg; niacin, 30 mg; biotin, 0.06 mg; iodine, 0.8 mg; copper, 10 mg; iron, 80 mg; selenium, 0.3 mg; manganese, 80 mg; and zinc, 80 mg. 2 Data analysed in technical duplicates.
Table 2. The proximate analysis of the studied wheat distillers’ dried grains with solubles (DDGS) 1.
Table 2. The proximate analysis of the studied wheat distillers’ dried grains with solubles (DDGS) 1.
Wheat DDGS
Dry Matter (g/kg)896.0
Gross Energy (MJ/kg DM)21.78
Crude Protein (g/kg DM)354.0
Fat (g/kg DM)49.8
Neutral Detergent Fibre (g/kg DM)493.3
Ash (g/kg)54.9
Total Non-Starch Polysaccharides (g/kg DM)234.5
1 Data analysed in technical duplicates.
Table 3. Analysis of xylanase (XYL), phytase (PHY), and protease (PRO) activity in the experimental diets 1.
Table 3. Analysis of xylanase (XYL), phytase (PHY), and protease (PRO) activity in the experimental diets 1.
DietsExpectedAnalysed
XYLPHYPROXYLPHYPRO
No exogenous enzyme added00003390
XYL2000002718327150
PHY05000174746132
PRO0040001423634142
XYL × PHY200050001636684241
XYL × PRO20000400021213554054
PHY × PRO050040001368993967
XYL × PHY × PRO2000500400013936163870
1 The diets consisted of 8 experimental treatments: (No exogenous enzyme added) the basal feed without XYL, PHY, or PRO supplementation; (XYL) the basal feed supplemented with XYL only; (PHY) the basal feed supplemented with PHY only; (PRO) the basal feed supplemented with PRO only; (XYL × PHY) the basal feed supplemented with XYL and PHY only; (XYL × PRO) the basal feed supplemented with XYL and PRO only; (PHY × PRO) the basal feed supplemented with PHY and PRO only; (XYL × PHY × PRO) the basal feed supplemented with XYL, PHY, and PRO.
Table 4. The effects of the experimental diets on apparent metabolisable energy (AME), dry matter (DM), nitrogen, fat, and neutral detergent fibre (NDF) ileal digestibility coefficients when fed to laying hens from 22 to 23 weeks of age (the AME data are based on the last 4 days of the excreta collection period; digestibility coefficients obtained on ileal digesta).
Table 4. The effects of the experimental diets on apparent metabolisable energy (AME), dry matter (DM), nitrogen, fat, and neutral detergent fibre (NDF) ileal digestibility coefficients when fed to laying hens from 22 to 23 weeks of age (the AME data are based on the last 4 days of the excreta collection period; digestibility coefficients obtained on ileal digesta).
Treatment FactorAME
(MJ/kg)		Digestibility Coefficients
DMNitrogenFatNDF
No exogenous enzyme added10.330.6400.6480.8940.117
XYL11.380.6360.6460.8870.114
PHY10.780.6220.6170.8910.117
PRO11.000.6230.6170.8490.120
XYL × PHY11.310.6300.6280.8680.117
XYL × PRO11.370.6650.6640.8950.128
PHY × PRO11.190.6310.6210.9030.126
XYL × PHY × PRO11.600.6790.7080.9160.155
SEM0.3790.02510.03200.02610.0234
XYL
-10.820.6290.6260.8840.120
+11.410.6520.6610.8920.123
PHY
-11.020.6410.6430.8810.120
+11.220.6410.6440.8950.123
PRO
-10.950.6320.6350.8850.111
+11.290.6500.6520.8910.132
SEM0.1890.01310.01610.01300.0165
Probabilities of statistical differences
XYL0.0350.1930.1200.6870.828
PHY0.4660.9990.9930.4590.770
PRO0.2170.3230.4280.7510.082
XYL × PHY0.6610.8080.5630.4910.966
XYL × PRO0.4560.2270.1710.2270.182
PHY × PRO0.9730.5210.2830.1840.263
XYL × PHY × PRO0.5980.9450.7680.8340.407
XYL—2000 U xylanase/kg diet; PHY—500 U phytase/kg diet; PRO—4000 U protease/kg diet; SEM—standard error of means; there is a statistically significant difference when p < 0.05.
Table 5. The effect of the experimental diets on daily feed intake (FI), daily body weight gain (WG), egg production, and feed conversion ratio (FCR) for egg production when fed to laying hens from 22 to 23 weeks of age (data are based on a 9-day feeding period).
Table 5. The effect of the experimental diets on daily feed intake (FI), daily body weight gain (WG), egg production, and feed conversion ratio (FCR) for egg production when fed to laying hens from 22 to 23 weeks of age (data are based on a 9-day feeding period).
Treatment factorWeight Gain
(g/hen/d)		FI
(g DM/hen/d)		Eggs
(hen/d)		Egg Mass
(g/hen/d)		FCR
(kg:kg)
No exogenous enzyme added1.779.20.7241.71.897
XYL1.482.50.7041.51.999
PHY3.582.90.7242.21.972
PRO2.183.90.7143.41.937
XYL × PHY2.383.50.7143.71.908
XYL × PRO4.680.20.6738.92.107
PHY × PRO7.883.40.7242.31.977
XYL × PHY × PRO3.687.50.7042.42.065
SEM1.393.070.0221.460.0934
XYL
-3.182.40.7242.41.946
+3.083.40.6941.62.020
PHY
-2.581.50.7041.41.985
+3.684.30.7142.61.980
PRO
-2.282.00.7142.31.944
+3.983.70.7041.82.021
SEM0.982.170.0161.030.0661
Probabilities of statistical differences
XYL0.9010.6270.1610.4600.270
PHY0.2540.1990.3290.2370.945
PRO0.1050.4320.4460.6140.249
XYL × PHY0.2180.5690.7430.1340.355
XYL × PRO0.5350.6880.5860.1680.414
PHY × PRO0.8760.8090.5860.9240.957
XYL × PHY × PRO0.4570.2330.7430.4910.754
XYL—2000 U xylanase/kg diet; PHY—500 U phytase/kg diet; PRO–4000 U protease/kg diet; SEM—standard error of means; there is a statistically significant difference when p < 0.05.
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MDPI and ACS Style

Whiting, I.M.; Rose, S.P.; Amerah, A.M.; Pirgozliev, V.R. Enzyme Supplementation to Diets Containing Wheat Distillers’ Dried Grains with Solubles (DDGS) When Fed to Laying Hens. Poultry 2025, 4, 22. https://doi.org/10.3390/poultry4020022

AMA Style

Whiting IM, Rose SP, Amerah AM, Pirgozliev VR. Enzyme Supplementation to Diets Containing Wheat Distillers’ Dried Grains with Solubles (DDGS) When Fed to Laying Hens. Poultry. 2025; 4(2):22. https://doi.org/10.3390/poultry4020022

Chicago/Turabian Style

Whiting, Isobel M., Stephen P. Rose, Ahmed M. Amerah, and Vasil R. Pirgozliev. 2025. "Enzyme Supplementation to Diets Containing Wheat Distillers’ Dried Grains with Solubles (DDGS) When Fed to Laying Hens" Poultry 4, no. 2: 22. https://doi.org/10.3390/poultry4020022

APA Style

Whiting, I. M., Rose, S. P., Amerah, A. M., & Pirgozliev, V. R. (2025). Enzyme Supplementation to Diets Containing Wheat Distillers’ Dried Grains with Solubles (DDGS) When Fed to Laying Hens. Poultry, 4(2), 22. https://doi.org/10.3390/poultry4020022

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