Animal Science

Effect of inclusion levels of dry distillery grains with solubles (DDGS) on productivity and egg quality of Hy-Line Brown^® laying hens

F. Castiblanco¹ 
http://orcid.org/0000-0002-1685-5367

P. E. Paz² 
http://orcid.org/0000-0002-9388-9579

M. Valdivié³ 
http://orcid.org/0000-0002-8858-0307

Y. Martínez^{1  *} 
http://orcid.org/0000-0003-2167-4904

¹ Centro de Investigación y Enseñanza Avícola, Departamento de Ciencia y Producción Agropecuaria, Escuela Agrícola Panamericana. Zamorano, Honduras

²Departamento de Ciencia y Producción Agropecuaria, Escuela Agrícola Panamericana, Zamorano, Honduras

³Centro Nacional para la Producción de Animales de Laboratorio Santiago de las Vegas, Rancho Boyeros, La Habana, Cuba

Abstract

The effect of four levels of inclusion of dry distillery grains with solubles on the productive performance and egg quality of Hy-line Brown^® laying hens was evaluated. A total of 140 animals were distributed in four treatments, according to a completely randomized design, for 10 weeks. Seven repetitions per treatment and five birds per repetition were established. The treatments consisted of a control diet and the inclusion of 10, 15 and 20 % of dry distillery grains with solubles. The inclusion of 0 and 10 % promoted (P <0.05) laying intensity and mass conversion with respect to 15 and 20 %. In addition, diets with dry distillery grains with solubles increased (P <0.05) food intake, nutrients and metabolizable energy, as well as shell thickness and yolk color (P <0.05), without notable changes (P> 0.05) for albumen height, Haugh unit and resistance to shell rupture. It is recommended the inclusion of 10 % of dry distillery grains with solubles in partial substitution of corn and soybean meal in diets intended for Hy-line Brown^® laying hens.

Key words: distillery byproducts; laying hens; productive indicator; egg external and internal quality

Currently, the demand for imported ingredients, such as corn and soybean, mainly, as well as the mobility restrictions due to COVID-19, have caused an increase of the price of these inputs, causing logistical problems for animal feed factories and poultry industry (^{USDA 2020}). Thus, it is necessary to explore new food sources, available to reduce the high production costs (^{Valdivié et al. 2020}). For some decades, dry distillery grains with soluble (DDGS) have been recommended as a food alternative available for ruminants and monogastric animals, with the aim of lowering production costs and reducing dependence on the use of corn and soybean meal (^{Abd El-Hack et al. 2015}). Due to the increase of ethanol production as a biofuel, the availability of DDGS has increased considerably (^{Zhu et al. 2018}), mainly in the United States, which produces more than 44 million metric tons per year (^{U.S Grains Council 2021}).

DDGS are rich in crude protein, available phosphorus, essential amino acids, and vitamins (^{Swiatkiewicz and Koreleski 2008}). Although DDGS are produced from other cereals, the feed industry prefers those that come from corn fermentation, because the levels of neutral detergent fiber (NDF) and acid detergent fiber (ADF) are relatively low and reduce problems related to feed digestibility (^{Szambelan et al. 2020}). ^{Jiang et al. (2013)} recommend the inclusion between 10 and 12 % of DDGS in the diet, without reducing the productivity and quality of the egg of laying hens. ^{Wu-Haan et al. (2010)} reported that the inclusion of 20% of DDGS in diets of laying hens did not cause adverse effects on the productive response and reduced the emission of NH₃ and H₂. ^{Shalash et al. (2010)} pointed out that the inclusion of 20 % of DDGS plus exogenous enzymes could partially replace the energy and protein ingredients in diets, without affecting productivity of laying hens.

Despite the fact that several studies have been developed with different levels of DDGS use in laying hens (^{El-Hack et al. 2019}), there are still contradictions, mainly related to the technology for ethanol production, which has a direct impact on the chemical composition of DDGS (^{Böttger and Südekum 2018}). Nowadays, the technology for ethanol production has managed to standardize the process for obtaining DDGS and reduce the variability of its chemical components. Furthermore, the fermentation process has substantially reduced starch concentration and non-enzymatic protein glycosylation, with the aim of increasing crude protein and amino acid availability (^{Iram et al. 2020}). It is necessary to continue the study of DDGS in diets for hens, to elucidate the optimal inclusion level and its effect on the main indicators in laying hens.

The objective of the study was to evaluate the effect of the inclusion of DDGS levels on the productive performance and egg quality of Hy-Line Brown^® laying hens.

Materials and Methods

Experimental location. This study was conducted at the Poultry Research and Teaching Center of the Escuela Agrícola Panamericana, in Zamorano, Honduras. This facility is located in Valle del Yeguare, San Antonio de Oriente municipality, Francisco Morazán department, 32 km from Tegucigalpa, Honduras. It has a height of 800 m o. s. l. and average temperature of 26 °C.

Animals, experimental design and treatments. A total of 140 Hy-Line Brown^® laying hens, 77 weeks old, were distributed according to a totally randomized design for 10 weeks, with four treatments, seven repetitions per treatment and five animals per repetition. Dietary treatments consisted of a control diet (T0) and the inclusion of 10 (T1), 15 (T2) and 20 % (T3) of DDGS.

DDGS were acquired from Alimento company, in Tegucigalpa, Honduras. The marketing company reported the following chemical composition: 81.32 % of dry matter, 9.33 MJ/kg of metabolizable energy, 28.13 % of crude protein, 7.5 % of crude fiber, 0.54 % of digestible lysine, 0.85% of digestible methionine+cystine and 0.75 % of digestible threonine. In addition, the content of ash (6.76 %), K (1.37%), Ca (0.1 %), Mg (0.42 %), Cu (7.67 mg/kg), Fe (97.67 mg/kg), Mn (24.67 mg/kg) and Zn (109 mg/kg), according to ^{AOAC 2001.11 (2006)} and P (0.60 %) by the colorimetry method with molybdenum blue, were determined in DDGS by triplicate, in the Soils laboratory of the University by Zamorano. Diets were formulated according to the nutritional requirements described in the manual of the used genetic line (table 1).

Table 1 Ingredients and nutritional contributions of the diets of laying hens (77-86 weeks) 

Ingredients, %	DDGS inclusion levels, %
	0	10	15	20
Corn meal	73.13	65.82	62.09	58.29
Soybean meal	12.53	8.81	6.86	4.98
DDGS	0.00	10.0	15.0	20.0
Mineral and vitamin premix^1	0.20	0.20	0.20	0.20
Sodium chloride	0.30	0.30	0.30	0.30
African palm oil	0.00	1.18	1.77	2.38
Wheat bran	0.39	0.00	0.00	0.00
Choline	0.05	0.05	0.05	0.05
DL-Methionine	0.30	0.30	0.31	0.31
L-Threonine	0.14	0.16	0.17	0.18
L-Lysin	0.29	0.36	0.40	0.43
Fine calcium carbonate	3.42	3.43	3.43	3.43
Thick calcium carbonate	7.99	7.99	8.00	8.00
Biofos	1.14	1.2	1.22	1.25
Mycofix plus 5.0	0.12	0.20	0.20	0.20
Nutritional contribution, %
ME, MJ/kg	11.40	11.40	11.40	11.40
Crude protein	14.22	14.22	14.22	14.22
Ca	4.44	4.44	4.44	4.44
Available P	0.35	0.35	0.35	0.35
Lysin	0.76	0.76	0.76	0.76
Methionine + cystine	0.72	0.72	0.72	0.72
Threonine	0.57	0.57	0.57	0.57

¹Mineral and vitamin premix: vitamin A, 1,000 IU/kg; vitamin D₃, 2,000 IU /kg; vitamin E, 30 IU/kg; vitamin K₃, 2.0 mg/kg; vitamin B₁, 1.0 mg/kg; vitamin B₂, 6.0 mg/kg; vitamin B₆, 3.5 mg/kg; vitamin B₁₂, 18 mg/kg; niacin, 60 mg/kg; pantothenic acid, 10 mg/kg; biotin, 10 mg/kg; folic acid, 0.75 mg/kg; choline, 250 mg/kg; iron, 50 mg/kg; copper, 10 mg/kg; zinc, 70 mg/kg; manganese, 70 mg/kg; selenium, 0.30 mg/kg; iodine, 1.0 mg/kg

DDGS: dry distillery grains with solubles

Experimental conditions. The laying hens were housed in a 400 m² commercial shed, in 61×36 cm cages, with ceiling fans and an artificial lighting system. Water was offered ad libitum in two nipple drinkers per cage and feed intake was restricted to 115 g/d/hen in linear feeders. Sixteen hours of light were provided each day, and no therapeutic veterinary care was used during the experimental stage. To achieve adequate adaptation to the new diets, a 7-day pre-experimental feeding phase was used, recommended by ^{Abd El-Hack et al. (2015)}.

Productive performance. To determine laying intensity (LI), total egg production/week/treatment was considered. One egg/d/housed hen was assumed to be 100 %. To determine egg weight (EW), 30 eggs were collected weekly per each treatment, between 8:30 a.m. and 9:30 a.m. Eggs were weighed on an OHAUS^® digital technical balance (New Jersey, USA), with an accuracy of ± 0.1 g. Mortality was determined by considering dead animals among those that started the experiment. Food intake (FI) and nutrients was determined three times a week, according to the offer and reject method. This means intake of metabolizable energy (MEI), crude protein (CPI), calcium (CaI), phosphorus (PI), lysine (LysI), methionine + cystine (Met + cysI), threonine (ThrI) and metabolizable energy (ME). Mass conversion (MC) and unfit eggs (UFE) were calculated with the following formulas:

$\mathrm{M}\mathrm{C}=\frac{\mathrm{F}\mathrm{o}\mathrm{o}\mathrm{d}\mathrm{}\mathrm{i}\mathrm{n}\mathrm{t}\mathrm{a}\mathrm{k}\mathrm{e}}{\mathrm{N}\mathrm{u}\mathrm{m}\mathrm{b}\mathrm{e}\mathrm{r}\mathrm{}\mathrm{o}\mathrm{f}\mathrm{}\mathrm{e}\mathrm{g}\mathrm{g}\mathrm{s}\mathrm{}\mathrm{x}\mathrm{}\mathrm{e}\mathrm{g}\mathrm{g}\mathrm{}\mathrm{w}\mathrm{e}\mathrm{i}\mathrm{g}\mathrm{h}\mathrm{t}}$

$\mathrm{U}\mathrm{F}\mathrm{E}=\frac{\mathrm{U}\mathrm{F}\mathrm{E}\mathrm{*}100}{\mathrm{G}\mathrm{o}\mathrm{o}\mathrm{d}\mathrm{}\mathrm{e}\mathrm{g}\mathrm{g}\mathrm{s}}$

Egg external and internal quality. At week 86, 30 eggs were collected per each experimental treatment. All were collected at the same time and transferred to the egg quality laboratory of the Research and Teaching Center of Zamorano Pan-American Agricultural School. Egg quality was analyzed on the same day of collection using an automatic TSS EggQuality analyzer (York, England) and Eggware v4x program. Resistance to rupture of the eggshell (middle pole) was measured with a QC-SPA^® resistance analyzer (York, England).

For analyzing egg shell thickness (EST) (middle pole), a QC-SPA^® micrometer screw (York, England) was used with a precision of ± 0.001 mm. For internal egg quality, albumen height (AH) was determined using a QHC^® height analyzer (York, England) with a precision of ± 0.01 mm. Haugh units were calculated with the formula:

$\mathrm{H}\mathrm{U}\mathrm{}=\mathrm{}100\mathrm{}\mathrm{*}\mathrm{}\mathrm{l}\mathrm{o}\mathrm{g}\mathrm{}(\mathrm{A}\mathrm{H}\mathrm{}+\mathrm{}1.7\mathrm{E}\mathrm{W}0.37\mathrm{}+\mathrm{}7.6)$

where:

HU	is Haugh unit
AH	is albumen height
EW	is egg weight

Yolk color (YC) was evaluated using a CCC^® electronic colorimeter (York, England), which considers the Roche scale of 15 colors.

Statistical analysis. Data was processed by analysis of variance (Anova) in a completely randomized design. In the necessary cases, ^{Duncan test (1955)} was used. Furthermore, yolk color was determined by the non-parametric ^{Kruskal and Wallis (1952)} test, with Bonferroni correction (^{Armstrong 2014}). Values of P<0.05 were taken to indicate significant differences. The SPSS 23.0.1.2014 program (^{SPSS Inc., Chicago, IL, USA}) was used for statistical analyzes.

Results and Discussion

The inclusion of 10 % of DDGS in the diets of laying hens (P <0.05) showed laying intensity (P < 0.001) and mass conversion (P < 0.001) similar to control, and both treatments, with 15 and 20 % of DDGS inclusion, indicated differences (P < 0.05). This treatment (10 % of DDGS) also improved (P < 0.001) egg weight by 1.85 g in relation to control, although the inclusion of 20 % of DDGS showed (P <0.001) the lowest values. Likewise, the inclusion of DDGS (10, 15 and 20 %) caused higher food intake, metabolizable energy and nutrients in relation to the diet without DDGS (P < 0.001). The percentage of unfit eggs did not change due to the effect of experimental diets (P = 0.709) (table 2).

Table 2 Effect of DDGS inclusion on the productive performance of Hy-line Brown^® laying hens (77-86 weeks) 

Items	DDGS inclusion levels, %				SE±	P Value
	0	10	15	20
LI, %	85.55^a	85.10^a	80.29^b	80.49^b	0.408	<0.001
EW, g	63.15^c	65.00^a	63.63^b	62.83^d	0.043	<0.001
FI, g/hen/d	108.92^b	110.33^a	109.75^a	109.79^a	0.114	<0.001
MEI, kcal/hen/d	1.24^b	1.26^a	1.25^a	1.25^a	0.0005	<0.001
CPI, g/hen/d	15.49^b	15.69^a	15.61^a	15.61^a	0.016	<0.001
CaI, g/hen/d	4.83^b	4.90^a	4.87^a	4.87^a	0.005	<0.001
Available PI, mg/hen/d	381.8^b	385.8^a	383.4^a	384.5^a	0.380	0.002
LysI, mg/hen/d	827.5^b	837.6^a	834.2^a	834.5^a	0.840	<0.001
Met+CysI, mg/hen/d	785.9^b	794.1^a	790.5^a	791.7^a	0.810	<0.001
ThrI, mg/hen/d	620.8^b	628.9^a	625.6^a	625.8^a	0.650	<0.001
MC, kg/kg	2.18^b	2.11^b	2.31^a	2.39^a	0.019	<0.001
UFE, %	1.62	1.41	1.82	1.96	0.175	0.709

^a,b,c,dMeans with different superscripts in the same line differ at P < 0.05

One of the objectives of this study was to verify whether the dietary use of corn DDGS with new biotechnological processes to correct nutrient variability and eliminate non-enzymatic protein glycosylation could have a better productive response in laying hens fed 10, 15 and 20 % of DDGS. In this sense, the inclusion of DDGS caused higher food intake and, as well as metabolizable energy and nutrients (table 2), which may be related to the increase of hedonic nutrients in DDGS, such as lipids and tryptophan, with respect to corn meal (^{FEDNA 2018}).

^{Lisnahan and Nahak (2020)} indicated that a higher concentration of tryptophan in the diet stimulated food intake, related to an increase of the precursor of serotonin and niacin. ^{Shin et al. (2015)} and ^{Mahrose et al. (2016)} indicated that levels of 10 and 16.5 % of DDGS in diets of laying hens caused an increase of food intake. Also, ^{Dinani et al. (2018)} mentioned that the high nutritional value of DDGS has a certain relationship with obtaining this ingredient, since once starch is fermented, nutrients of corn grain are concentrated in large quantities in the DDGS particles. According to ^{Pottgüter (2015)}, the high intake of amino acids is due to their high content and bioavailability in dry distillery grains with solubles (DDGS), and their levels vary depending on the color tone of the product, this being an indicator of its quality. Regarding methionine and cystine intake, ^{El-Sheikh and Salama (2020)} mention that DDGS are rich sources of sulfur amino acids, when compared to protein foods such as soybean meal. A higher intake of DDGS increases, in turn, the intake of these amino acids. ^{Mutucumarana et al. (2014)} also report that phosphorus intake increases, due to its high presence in DDGS. According to ^{Abd El-Hack et al. (2018)}, phosphorus is highly available and with lower proportions of phytate, which is related to fermentation processes to which this ingredient is subjected.

Similar results in laying intensity and mass conversion were obtained by ^{Jiang et al. (2013)} and ^{Abd El-Hack et al. (2015)}, who recommended the inclusion between 10 and 12 % of DDGS in the diet. Also, ^{Roberson et al. (2005)} informed that the inclusion of 15 % of DDGS did not reduce the response in laying hens. ^{Wu-Haan et al. (2010)} and ^{Rodríguez et al. (2016)} reported no productive changes, when they used up to 20 % of DDGS in diets of laying hens. ^{Shalash et al. (2010)} indicated that the use of DDGS (20%) plus exogenous enzymes in the diets, did not affect laying intensity and mass conversion of laying hens. ^{Masa´deh et al. (2011)} stated that the inclusion of up to 25 % of DDGS in diets for laying hen did not affect egg production and mass conversion.

The variability of results with the use of DDGS in laying hens is due to the process of obtaining this by-product in ethanol production (^{Böttger and Südekum 2018}). Authors such as ^{Cromwell et al. (1993)}, ^{Spiehs et al. (2002)}, ^{Belyea et al. (2006)} and ^{Liu (2009)} reported ranges for crude protein (25.8 to 31.7 %), oil (9.1 to 14.1%), ash (3.7 to 8.1 %), NDF (33.1 to 43.9%), lysine (0.48 to 1.15% ), methionine (0.49 to 0.76 %) and threonine (0.99 to 1.28 %), so it is necessary to determine the chemical composition of DDGS prior to the productive study.

Results of table 2 showed that the inclusion of 15 and 20 % of DDGS in substitution of 11.04 and 14.84% of corn meal and 5.67 and 7.55 % of soybean meal, respectively, decreased the productive response of hens, although the experimental diets supplied essential and limiting amino acids, such as lysine, methionine + cystine, and threonine. Other essential and non-essential amino acids could be out of balance with the increased inclusion of DDGS. Additionally, the increased utilization of DDGS (up to 20%) and non-starch fibrous compounds probably reduced nutrient digestibility. ^{Abd El-Hack et al. (2015)} reported that the inclusion of up to 18 % of DDGS reduced the digestibility coefficient of dry matter (2.06 %), organic matter (4.57 %), crude protein (10.14 %), crude fat (9.84 %) and crude fiber (18.58 %).

In this study, despite the variability of results, laying intensity in the experimental treatments (80.40 to 85.55 %) was higher than that proposed by the ^{Hy-Line Brown management guide (2018)} for hens aged between 77-86 weeks (77 to 74 %).

Treatment with 10 % of DDGS showed the highest egg weight. This may be justified by a higher intake of sulfur amino acids with respect to control treatment and by a lower intake of crude fiber, related to treatments with 15 and 20 % of DDGS (table 2).

Methionine is known to be the essential and limiting amino acid, related to egg weight (^{Martínez et al. 2017}). ^{Faria et al. (2003)} found that an increment of 1.14 % of the intake of methionine + cystine in laying hens promoted an increase of egg weight, similar to that recorded in this study, in which there was an increase in the intake of these sulfur amino acids (AA) by 1.26 %. The decrease of egg weight in treatments with 15 and 20 % DDGS could be also related to the reduction of digestibility of the amino acids of this ingredient, especially of the unbalanced AA in the diets. According to ^{Pottgüter (2015)} and ^{Dinani et al. (2018)}, high levels of DDGS in diets reduce digestibility of nutrients, especially proteins and amino acids, mainly due to the concentration of crude fiber and the possible non-enzymatic glycosylation of proteins. Similar results were reported by ^{Masa´deh et al. (2011)}, who state that egg weight tends to decrease as DDGS increase in the diet. However, studies of ^{Swiatkiewicz et al. (2013)} and ^{Yildiz et al. (2018)} did not find significant changes in egg weight, when they used up to 30 % of DDGS in diets for laying hens, so different responses of egg weight to the increase of DDGS in the diet are observed.

From the statistical point of view, mass conversion showed similar results for control diet and treatment with 10 % of DDGS. Higher levels of DDGS inclusion increased this productive indicator (table 2). ^{Rodríguez et al. (2016)} reported that mass conversion was not affected with levels of 10 and 20 % of DDGS in diets intended for layers. However, ^{Saeed et al. (2017)} pointed out the negative effect of this productive parameter (mass conversion) when using 20 % of inclusion of DDGS in the diet of laying hens, which is associated with a decrease of weight and production of eggs. According to ^{Elshikha et al. (2018)}, mass conversion increases with higher inclusion of DDGS in diets, due to the decrease of egg production and the increase of food intake. The variability of the chemical composition of DDGS, due to biotechnological processes, is the main cause of variation in food intake and in production and weight of eggs of laying hens, important to determine this productive indicator (mass conversion).

It is known that eggs classified as unfit generate considerable economic losses in productive units, as they are not marketed. These problems, according to ^{England and Ruhnke (2020)}, are due to the fact that the calcification process of laying hens is less efficient, with greater emphasis on old hens due to the decrease of sex hormones (mainly estrogens). ^{Mazzuco and Bertechini (2014)} ensure that the incidence of unfit eggs is directly related to the quality of the shell, in combination with other factors.

Previous results show that the use of up to 20% of DDGS in the diet did not affect the inclusion of calcium ions into the shell. The levels of consumed crude fiber had an influence on the absorption of this mineral, because, according to ^{Savón et al. (2007)}, high levels of crude fiber in poultry diets increase mineral restriction, and affect the absorption of minerals such as Ca and Mg, as well as eggshell quality.

Table 3 shows the effect of the inclusion of 0, 10, 15 and 20 % of DDGS on external and internal egg quality at 87 weeks of age of hens. The inclusion of DDGS significantly increased egg weight (P = 0.03), shell thickness (P<0.001) and yolk color (P<0.001). However, albumen height (P = 0.100), Haugh unit (P = 0.221) and resistance to rupture (P=0.149) of the shell were not modified due to the experimental treatments (P > 0.05).

Table 3 Effect of the inclusion of dry distillery grains with solubles on external and internal egg quality of Hy-line Brown^® laying hens (86 weeks) 

Egg quality	DDGS inclusion levels (%)				SE±	P Value
	0	10	15	20
EW (g)	60.03^b	63.56^a	63.51^a	62.71^a	0.390	0.003
AH (mm)	10.37	10.96	11.14	10.79	0.114	0.100
HU	100.14	102.32	103.1	101.74	0.518	0.221
RR (kgF/cm^2)	4340.00	4425.10	4368.00	4360.68	65.91	0.149
ST (mm)	0.33^b	0.36^a	0.38^a	0.37^a	0.004	<0.001
YC	4.00^b (0.050)	5.00^a (0.052)	5.00^a (0.051)	5.00^a (0.051)		<0.001

^a,b Means with different superscripts in the same line differ at P < 0.05

( ) standard deviation

In this experiment, it could be seen that the inclusion of DDGS had a positive influence on shell thickness, a result that coincides with that reported by ^{Sedmake et al. (2018)}, who indicated that the inclusion of 20% of DDGS increased shell thickness, due to the high concentration of orthophosphate (available P) in this food. ^{Morales et al. (2016)} mentioned that, in the biotechnological process, DGGS triples the mineral composition with respect to corn meal, which could have an impact on mineral metabolism and shell thickness. According to ^{Sun and Kim (2020)}, increasing phytic P intake reduces Ca absorption and, therefore, egg shell thickness in laying hens, due to the imbalance in Ca: P ratio. Values of resistance to rupture also coincide with those reported by ^{Shin et al. (2016)}, who mentioned that DDGS inclusion had no effect on this external quality parameter. ^{Nasri et al. (2020)} indicated that the optimal quality of the shell determines egg durability on the shelf and its greater commercialization.

The current study also evidenced greater intensity in yolk color in the treatments with DDGS (table 3). The golden color of DDGS is due to the carotenoids found in yellow corn, used for ethanol production (^{Abd El-Hack et al. 2015}). ^{Shin et al. (2016)} mentioned that, due to the biotechnological process, high concentrations of lutein and zeaxanthin in DDGS cause greater intensity in egg yolk color, when this product is used up to 10 % in the diet. This agrees reports of ^{Cortes-Cuevas et al. (2015)} and ^{Shin et al. (2015)}, who observed an increase of yolk color with the use of DDGS in the diet (up to 15 %). Currently, in many countries, yolk color constitutes one of the main parameters for consumers to decide to buy, who prefer a more pigmented yolk (^{Martínez et al. 2021}).

Albumen height and Haugh unit did not change due to DDGS diets (table 3). This demonstrates that the inclusion of up to 30 % of DDGS did not affect protein synthesis in albumen, since the amount of white depends on the amino acid balance provided by the protein in the diet. Lysine or methionine deficiency reduces albumen weight, and decreases the concentration of all free amino acids (^{Sun et al. 2019}). Similar results were informed by ^{Yildiz et al. (2018)}, who used feedstuff with 0, 10, 20 and 30 % of DDGS, without observing changes in these parameters. Data of albumen height agree with ^{Abd El-Hack et al. (2015)}, who indicated that the inclusion of DDGS at levels up to 18% did not affect this variable. Albumen height and Haugh unit are considered as the most important internal quality indicators, related to egg freshness (^{Narushin et al. 2021}).

Conclusions

The evaluated DDGS, which presents 28.13 % of crude protein, 9.33 MJ ME/kg, 7.5 % of crude fiber, 0.54 % of digestible lysine, 0.85 % of digestible methionine + cystine and 0.75 % of digestible threonine, can be efficiently used with a 10 % level in diets for 77-week-old Hy-Line Brown^® laying hens. This allows the animals to express their maximum potential for egg production and to produce heavier eggs, with thicker shells, higher yolk pigmentation and the same mass conversion compared to control treatment. Meanwhile, the inclusion of 15 and 20 % of this DDGS reduced egg production of hens and damaged mass conversion and egg weight.

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Received: September 02, 2021; Accepted: October 08, 2021

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Conflict of interests: The authors declare that there is no conflict of interest among them.

Contribution of authors: F. Castiblanco: Conduction of the experiment, data analysis and paper writing. P. E. Paz: Data analysis. M. Valdivié: Data analysis and paper writing. Y. Martínez: Original idea, conducting the experiment, research design