Effects of selected feed additives on the performance

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Effects of selected feed additives on the performance of laying hens given a diet rich in maize dried distiller's grains with solubles (DDGS) a

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S. Świątkiewicz , A. Arczewska-Włosek , J. Krawczyk , M. Puchała & D. Józefiak

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Department of Animal Nutrition and Feed Science , National Research Institute of Animal Production , Balice , Poland b

Department of Animal Nutrition and Feed Management , Poznań University of Life Sciences , Poznań , Poland Accepted author version posted online: 27 Apr 2013.Published online: 04 Jul 2013.

To cite this article: S. witkiewicz , A. Arczewska-Wosek , J. Krawczyk , M. Puchaa & D. Jzefiak (2013) Effects of selected feed additives on the performance of laying hens given a diet rich in maize dried distiller's grains with solubles (DDGS), British Poultry Science, 54:4, 478-485, DOI: 10.1080/00071668.2013.797563 To link to this article: http://dx.doi.org/10.1080/00071668.2013.797563

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British Poultry Science, 2013 Vol. 54, No. 4, 478–485, http://dx.doi.org/10.1080/00071668.2013.797563

Effects of selected feed additives on the performance of laying hens given a diet rich in maize dried distiller’s grains with solubles (DDGS) S. ŚWIĄTKIEWICZ, A. ARCZEWSKA-WŁOSEK, J. KRAWCZYK, M. PUCHAŁA AND D. JÓZEFIAK1

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Department of Animal Nutrition and Feed Science, National Research Institute of Animal Production, Balice, Poland, and 1Department of Animal Nutrition and Feed Management, Poznań University of Life Sciences, Poznań, Poland

Abstract 1. A total of 192 ISA Brown hens were given diets containing a high concentration of maize dried distiller’s grains with solubles (DDGS) and the effect of selected feed additives on laying performance and egg quality was determined. 2. Birds were allocated to 8 treatment groups with 12 replicates (cages) of two hens and were given, from week 26 to 55, iso-caloric and iso-nitrogenous experimental diets with or without a high concentration of DDGS (200 g/kg). The diet containing DDGS was not supplemented or supplemented with enzymes (xylanase and phytase), sodium butyrate, probiotic bacteria (Lactobacillus salivarius) and a mixture of herbal extracts (Taraxaci siccum, Urticae siccum and Salviae siccum), inulin or chitosan. 3. The inclusion of DDGS in the diet had no effect on number of eggs produced, total egg mass, mean egg weight, feed intake or feed conversion ratio. Egg and eggshell quality parameters were also unaffected by dietary DDGS. The yolk colour score (points in Roche scale) was significantly increased by DDGS inclusion. DDGS in the diet caused some changes in the yolk lipid profile that were rather unfavourable from a dietary perspective (an increase of cholesterol content, and PUFA n-6/PUFA n-3 ratio). 4. During the experimental period (26–55 weeks of age) supplementation of the diet containing a high concentration of DDGS with enzymes, inulin as well as chitosan, increased number of eggs produced and daily egg mass. In older hens (50 weeks of age) inulin positively affected eggshell quality parameters, i.e. shell percentage, thickness and density. Diet supplementation with herb extracts, inulin or chitosan, decreased the content of cholesterol in yolks. 5. The results of this study suggest that DDGS may be incorporated up to a concentration of 200 g/kg in the diet of laying hens without any negative effects on egg performance. Moreover, supplementation of xylanase and phytase, as well as inulin and chitosan, can positively affect the performance of layers given diets with a high concentration of DDGS.

INTRODUCTION Biofuels produced from grains are a renewable source of energy and they have a lower CO2 emission when burned than conventional fossil fuels. Thus the European Union supports biofuel production in order to diversify energy supplies, reduce greenhouse gas emission and dependency on oil and to create additional employment in rural areas. Distiller’s dried grains with solubles (DDGS) is a by-product of the ethanol industry created in the fermentation process of cereal

grains, mainly maize, and can be defined as the product obtained after the removal of ethyl alcohol by distillation from the yeast fermentation of a grain by condensing and drying at least 75% of the resultant whole stillage by methods employed in the grain distilling industry (AAFCO, 2002). During fermentation, approximately equal portions of ethanol, DDGS and CO2 are formed (Lumpkins et al., 2005). DDGS, largely as a by-product of the beverage industry, has been used as livestock feed for many decades, but due to the relatively high

Correspondence to: S. Świątkiewicz, Department of Animal Nutrition and Feed Science, National Research Institute of Animal Production, 32-083 Balice, Poland. E-mail: [email protected] Accepted for publication 12 February 2013.

© 2013 British Poultry Science Ltd

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DDGS DIETS FOR LAYERS

concentrations of fibre and variability in bioavailability of lysine as well as other nutrients, it has been traditionally given mainly to ruminants. In the early studies on laying hens, it was shown that DDGS was a valuable source of protein and sulphur amino acids and could be used at 50–200 g/kg inclusion concentrations in diets, even as a source of one-third of protein supply, without affecting performance (Matterson et al., 1966; Harms et al., 1969; Jensen et al., 1974) and had a positive effect on some egg quality parameters (Jensen et al., 1978). An increase in fuel ethanol production has resulted in an enlarged quantity of DDGS entering the feed market in recent years. The high nutritional quality of DDGS obtained from the modern ethanol industry is related to gentler drying conditions than in previous decades and allows the use of this feed material on a largescale in livestock and poultry nutrition, as the most effective and environmentally friendly way of using DDGS (Świątkiewicz and Koreleski, 2008). In an earlier study, it was concluded that maize DDGS is a useful feed ingredient for laying hens (Świątkiewicz and Koreleski, 2006). Moreover, results of a recent experiment with layers suggested that an addition of DDGS to diets could have a positive influence on nitrogen and phosphorus balance by increasing retention and decreasing faecal excretion of these elements (Masa’deh et al., 2011). During fermentation, cereal starch is converted to ethanol and CO2 and the concentration of remaining nutrients in DDGS increases by 2–3 times, so DDGS contains a high concentration of non-starch polysaccharides (NSPs). Therefore, as observed in some studies, reducing performance in hens given a diet with a high content of DDGS could be caused by a decrease in nutrient and energy utilisation. In our previous experiment, dietary concentrations of up to 150 g DDGS/kg diet had no negative effect on laying performance and egg quality, whereas a higher dietary concentration (200 g/kg) had a negative effect on laying rate and feed conversion (Świątkiewicz and Koreleski, 2006). Similar conclusions on the use of DDGS in layer diets were made by other authors (Lumpkins et al., 2005; Roberson et al., 2005; Shalash et al., 2010; Niemiec et al., 2012). Due to increasing prices of soybean protein, increasing concentrations of DDGS in poultry diets can be economically justifiable, especially in countries that depend on soybean meal import. In the literature, there is limited information on the effects of high concentrations of DDGS on egg production parameters. Therefore, the objective of the experiment was to determine the possibility of improving the performance of hens given a diet with a high concentration of maize DDGS (200 g/ kg) by using selected feed additives with potential

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positive effects on nutrient utilisation, i.e. enzymes (xylanase and phytase), sodium butyrate, probiotic bacteria (Lactobacillus salivarius), a mixture of herbal extracts (Taraxaci siccum, Urticae siccum and Salviae siccum), inulin or chitosan.

MATERIALS AND METHODS Birds and experimental diets A total of 192, 18-week-old ISA Brown hens, obtained from a commercial source, were placed in a poultry house, in cages (two birds per cage) having wire-mesh floor, and under controlled climate conditions. The cage dimensions were 30 cm × 120 cm × 50 cm, equating to 3600 cm2 total floor space. During the pre-experimental period (up to 26 weeks of age), a commercial laying hen diet (170 g/kg crude protein, 11.6 MJ/kg AMEN, 37.0 g/kg calcium and 3.8 g/kg available phosphorus) was offered ad libitum. At week 26, the hens were randomly assigned to one of 8 treatments, each comprising 12 replicates (cages with two hens in each) and given experimental diets until week 65. During the experiment, the hens had free access to feed and water, and were exposed to a 14 L:10 D lighting schedule, with a light intensity of 10 lux. The local Krakow Ethics Committee for Experiments with Animals approved all the experimental procedures relating to the use of live animals. Prior to formulating the experimental diets, it was determined (AOAC, 2000) that the DDGS used contained 269 g/kg crude protein, 11 g/kg crude fat, 5.50 g/kg crude fibre, 4.20 g/kg crude ash, 0.740 g/kg Lys, 0.620 g/kg Met, 1.04 g/kg Tre, 0.295 g/kg Try, 0.050 g/kg Ca, 0.826 g/kg P and 0.243 g/kg Na, respectively. Ingredient and nutrient composition of the basal diets used in the experiment is shown in Table 1. The diets without (Control group I) or with maize DDGS (200 g/ kg), were iso-caloric and iso-nitrogenous, and were formulated to meet or exceed nutrient recommendations (NRC, 1994). The diet with DDGS was not supplemented (group II) or supplemented with feed enzymes (Ronozyme WX with endo-1,4-β-xylanase activity of 1000 FXU/g and Ronozyme NP with phytase activity of 10 000 FYT/g, each preparation was added to the diet in the amount of 200 mg/kg, DSM Nutritional Products, group III), sodium butyrate (700 mg/ kg, GUSTOR XXI B 70, group IV), probiotic bacteria (L. salivarius, 108 cfu/kg, Institute of Agricultural and Food Biotechnology, group V), mixture of herbal (T. siccum, U. siccum and S. siccum) extracts (250 mg of each extract/kg, group VI), inulin (5 g/kg, BENEO™ IPS, Orafti, Belgium, group VII) or chitosan (3 ml/kg, Chimet-pasz, Gumitex Poli-Farm, Poland, group VIII). The nutrient content of the diets was

S. ŚWIĄTKIEWICZ ET AL.

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Table 1.

Composition and nutrient content of diets used in the experiment, g/kg

Item

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Maize Wheat Soybean meal DDGS Rapeseed oil Limestone Dicalcium phosphate Sodium chloride DL-Methionine L-Lysine HCl Vitamin-mineral premix1 Metabolisable energy (MJ/kg)2 Crude protein3 Lys3 Met3 Thr3 Ca3 Total P3 Digestible P3 Na3 DEB (mEq)4

Control diet (treatment I)

Experimental diets (treatments II– VIII)

360.0 255.6 245.0 – 24.0 90.0 16.0 3.0 1.4 – 5.0

290.0 201.0 165.0 200.0 28.0 92.0 13.0 3.0 1.1 1.9 5.0

11.6

11.6

175 8.6 4.1 5.8 37.0 6.0 3.8 1.5 175

175 8.6 4.1 6.0 37.0 6.0 3.8 1.8 185

1 The premix supplied per 1 kg of diet: retinyl acetate, 3.45 mg; cholecalciferol, 0.075 mg; DL-alpha-tocopheryl acetate, 20 mg; menadione sodium bisulphite, 2 mg; thiamine mononitrate, 1 mg; riboflavin, 4 mg; pyridoxine, 2 mg; cyanocobalamin, 0.015 mg; niacin, 25 mg; calcium pantothenate, 8 mg; folic acid, 0.5 mg; biotin, 50 µg; choline-HCl, 250 mg; Mn, 100 mg; Zn, 50 mg; Fe, 50 mg; Cu, 8 mg; I, 0.8 mg; Se, 0.2 mg; Co, 0.2 mg; 2 Calculated according to European Table (Janssen, 1989) as a sum of ME content of components; 3 Calculated according to the chemical composition of feed components; 4 Dietary electrolyte balance.

calculated according to the chemical composition of raw feed materials, metabolisable energy value– according to equations from European Tables (Janssen, 1989).

the time of eggshell fracture. At week 60 of age, one egg from each hen was collected to determine the fatty acid and cholesterol content of yolk lipids. The measurements were done directly after the eggs were collected. Chemical analyses The chemical composition of DDGS and other feed components was determined by conventional methods (AOAC, 2000). Amino acids were analysed in acid hydrolysates, after initial peroxidation of sulphur amino acids, in colour reaction with ninhydrin, using a Beckman-System Gold 126 AA automatic analyser. Calcium content by flame atomic absorption spectrophotometry and phosphorus content by calorimetric method were determined (AOAC, 2000). Cholesterol concentrations were determined in yolks by gas chromatography (Zhang et al., 1999). The fatty acid profile of the diets and yolks was determined on a VARIAN 3400 CX gas chromatograph, using helium as a carrier gas, and a 105 m Rtx 2330 column. Injector temperature was 200°C and detector temperature was 240°C. Samples were prepared according to Folch (1957) using methylation with BF3/methanol. Statistical analysis All data were subjected to statistical analysis using one-way ANOVA. When significant differences in treatment means were detected by ANOVA (Ftest), Duncan’s multiple range test was applied to the individual means. Differences were considered significant at P < 0.05. All statistical analyses were performed with Statistica 5.0 PL software (Statsoft, Inc., Tulsa, OK, USA).

RESULTS Laying performance

Sample collection During the experiment feed intake and number and weight of laid eggs were registered and laying performance, daily egg mass, daily feed intake and feed consumption per 1 kg of eggs and per 1 egg were calculated. The experimental period from 26 to 55 weeks of age was also divided in two phases, the first phase being from 26 to 39 weeks and the second from 40 to 55 weeks of age. At week 32 and week 50, one egg from each hen was collected to determine the egg quality, using semi-automated egg quality equipment (QCM+, Technical Services and Supplies (TSS), York, UK). Another egg was collected for measurements of shell breaking strength using Instron 5542 apparatus, equipped with a 500 Newton load cell. The eggs were compressed at a constant cross-head speed of 10 mm/ min and the breaking strength was determined at

The mean egg production, averaged across all dietary treatments, during the entire experimental period (26–55 weeks of hen age) was 92.0%; egg weight, 61.5 g; daily egg mass, 56.6 g/hen; daily feed consumption, 115 g/hen; feed utilisation, 124 g of feed consumed/1 egg and 2.03 kg of feed/1 kg of eggs (Table 2). During the entire experimental period, no dead hens were registered in any treatment. The inclusion of 200 g/kg DDGS to the diet had no significant effect on laying performance as compared to the control group, neither in the first nor in the second part of the laying phase (P > 0.05). During the experimental period some of the used feed additives had a beneficial effect on the performance indices of hens given the experimental diets (Table 2). Thus, the addition of feed enzymes significantly (P < 0.05) increased the

DDGS DIETS FOR LAYERS

Table 2.

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Effect of maize DDGS and experimental additives on laying performance Treatment

Item

I

II

III

0.923 54.9a 50.5 113 122 2.05

0.954 58.2b 61.1 116 122 1.99

0.945 56.5ab 59.8 115 122 2.04

0.929 56.7ab 61.0 115 124 2.04

Second phase of laying cycle (40–55 weeks of age) 0.895a Number of eggs produced/hen per d 0.907ab Daily mass of eggs, g per hen 56.9 55.9 Egg weight, g 62.8 62.4 Daily feed intake, g per hen 114 113 Feed, kg per 1 egg 126 126 Feed, kg per 1 kg of eggs 2.00 2.02

0.913b 56.8 63.3 115 126 2.03

0.905ab 55.9 61.8 114 126 2.04

Entire experimental period (26–55 weeks of age) Number of eggs produced/hen per d 0.914ab Daily mass of eggs, g per hen 56.4ab Egg weight, g 61.7 Daily feed intake, g per hen 114 Feed, kg per 1 egg 124 Feed, kg per 1 kg of eggs 2.02

0.928b 57.4b 61.8 116 124 2.01

0.919ab 56.1ab 61.1 115 125 2.04

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First phase of laying cycle (26–39 weeks of age) Number of eggs produced/hen per d 0.927 Daily mass of eggs, g per hen 55.6ab Egg weight, g 60.0 Daily feed intake, g per hen 114 Feed, kg per 1 egg 123 Feed, kg per 1 kg of eggs 2.05

0.905a 55.5a 61.4 113 125 2.03

IV

V

VI

VII

VIII

SEM

0.939 56.0ab 60.0 115 123 2.05

0.937 55.7ab 59.5 116 124 2.09

0.955 57.8ab 60.5 114 119 1.98

0.913b 57.3 62.8 116 127 2.02

0.907ab 55.9 61.7 113 125 2.03

0.919b 57.5 62.6 116 126 2.01

0.918b 57.4 62.1 115 125 2.01

000.2 0.179 0.116 0.310 0.312 0.055

0.918ab 57.1ab 62.2 115 126 2.03

0.916ab 56.0ab 61.1 114 124 2.04

0.925b 56.8ab 61.4 116 125 2.04

0.931b 57.3b 61.5 115 123 2.00

0.002 0.177 0.105 0.297 0.296 0.006

0.003 0.289 0.189 0.599 0.698 0.013

a,b Mean values within the same row sharing a common superscript letter are not statistically different at P < 0.05. Treatments: I – control diet, II–VIII – experimental diets containing 200 g/kg DDGS; II – not supplemented, III – supplemented with feed enzymes (xylanase + phytase), IV – supplemented with sodium butyrate, V – supplemented with probiotic bacteria, VI – supplemented with herb extracts mixture, VII – supplemented with inulin, VIII – supplemented with chitosan.

daily mass of eggs (group II vs group III) in the first phase of the laying cycle. Diet supplementation with enzymes, inulin or chitosan significantly (P < 0.05) improved the number of eggs produced in the second phase of the laying cycle (groups III, VII and VIII vs group II). Also, in the entire experimental period laying performance was beneficially affected when feed enzymes, inulin or chitosan, were added to the high DDGS diet (P < 0.05). Egg quality indices Most of the internal egg and eggshell quality parameters were not affected by dietary DDGS (P > 0.05), neither at 32 nor at 50 weeks of age (Table 3). Only the yolk colour score (points in Roche scale) was significantly increased by DDGS inclusion (P < 0.05) (Table 3). In most cases supplementation of the diet containing a high concentration of DDGS with experimental additives had no effect on internal egg and eggshell quality parameters; however, in older hens (50 weeks of age) inulin positively affected eggshell percentage, thickness and density (P < 0.05). Fatty acid and cholesterol contents in yolk lipids Dietary inclusion of a high concentration of DDGS significantly (P < 0.05) increased C18:2 n-6, C18:3n-3, C20:4 n-6, total PUFA, PUFA n-6 and

cholesterol concentrations and PUFA n-6/PUFA n-3 ratio. However, it decreased C18:1 and MUFA concentrations in yolk egg lipids as compared to hens given a control diet without DDGS (Tables 4 and 5). There were no statistically significant effects of a high DDGS diet on C16:0, C18:0, EPA, DHA, SFA, UFA and PUFA n-3 contents (P > 0.05). Experimental additives had no significant influence on the fatty acid profile of yolk lipids (P > 0.05), but diet supplementation with herb extracts, inulin as well as chitosan, decreased the content of cholesterol in yolks (P < 0.05) (Tables 4 and 5).

DISCUSSION The effects of a high dietary concentration of DDGS In this study, laying performance and feed conversion of hens given a diet with a high concentration of DDGS were very similar to production parameters observed in the control group. This lack of effect of high dietary DDGS can indicate that the experimental diets were well balanced and no nutritional deficiency existed. In our earlier study it was found that dietary concentrations up to 150 g/kg DDGS had no negative effect on laying performance, egg quality and flavour and taste of boiled eggs, whereas a 200 g/kg concentration of DDGS in the diet impaired laying rate and feed

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Table 3.

Effect of maize DDGS and experimental additives on internal and external quality of eggs Treatment

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Item

I

II

III

IV

V

VI

VII

VIII

SEM

32 weeks of age Albumen height, mm Haugh units Relative yolk weight, % Yolk colour, points in Roche scale Eggshell, % Eggshell thickness, μm Eggshell density, mg/cm2 Eggshell breaking strength, N

7.68 86.7 25.1 2.46a 11.0 390 88.2 34.8

7.66 86.9 24.8 3.33b 11.3 396 90.2 35.7

7.70 86.1 24.5 3.41b 11.4 408 92.2 37.0

7.41 84.7 24.6 3.29b 11.1 386 88.5 33.8

7.13 83.3 24.6 3.33b 11.0 390 88.1 34.2

7.04 82.8 24.5 3.46b 11.5 400 91.4 37.1

7.15 83.2 24.7 3.58b 11.5 395 89.9 36.5

7.46 85.1 25.0 3.50b 11.2 387 89.1 36.0

0.073 0.441 0.122 0.039 0.046 1.83 0.447 0.460

50 weeks of age Albumen height, mm Haugh units Relative yolk weight, % Yolk colour, points in Roche scale Eggshell, % Eggshell thickness, μm Eggshell density, mg/cm2 Eggshell breaking strength, N

6.41 73.5 26.9 1.83a 10.7ab 393ab 86.1ab 32.0

6.42 73.6 27.0 3.38b 10.5a 383a 83.3a 30.2

6.70 75.9 26.1 3.63b 10.7ab 385a 84.4a 31.1

6.65 75.3 27.0 3.63b 10.8ab 392ab 85.4ab 32.3

6.42 72.2 26.9 3.50b 10.7ab 392ab 85.7ab 30.8

6.52 73.5 26.6 3.33b 10.8ab 399ab 87.3ab 30.8

6.30 72.0 26.5 3.37b 11.1b 411b 89.3b 32.8

6.41 73.9 26.6 3.71b 11.0ab 398ab 87.9ab 31.9

0.064 0.414 0.123 0.050 0.047 1.98 0.427 0.434

a,b Mean values within the same row sharing a common superscript letter are not statistically different at P < 0.05. Treatments: I – control diet, II–VIII – experimental diets containing 200 g/kg DDGS; II – not supplemented, III – supplemented with feed enzymes (xylanase + phytase), IV – supplemented with sodium butyrate, V – supplemented with probiotic bacteria, VI – supplemented with herb extracts mixture, VII – supplemented with inulin, VIII – supplemented with chitosan.

Table 4. Effect of maize DDGS and experimental additives on concentrations of selected fatty acids and cholesterol in egg yolk lipids, % Treatment I II III IV V VI VII VIII SEM

C16:0

C18:0

C18:1

31.8 31.6 31.3 32.0 30.7 32.0 31.9 32.0 0.149

6.77 6.80 6.94 6.90 6.89 6.62 6.78 6.86 0.041

43.6b 39.2a 39.8a 38.8a 39.9a 39.3a 39.9a 39.5a 0.238

C18:2

n-6

9.67a 14.7b 15.4b 15.4b 15.5b 15.1b 15.0b 15.0b 0.285

C18:3

n-3

0.837a 0.908b 0.907b 0.929b 0.935b 0.911b 0.905b 0.912b 0.012

C20:4

n-6

C20:5 (EPA)

C22:6 (DHA)

Cholesterol (mg/g of yolk)

0.02 0.01 0.00 0.00 0.00 0.00 0.00 0.00 0.00

0.941 0.924 0.948 0.960 0.950 0.944 0.929 0.925 0.023

10.9a 12.4d 12.0cd 12.0cd 12.0cd 11.3ab 11.5bc 11.5bc 0.085

1.55a 1.89b 1.83b 1.74b 1.76b 1.72b 1.80b 1.77b 0.028

a,b Within a column, values not sharing a common superscript letter are significantly different (P ≤ 0.05). Treatments: I – control diet, II–VIII – experimental diets containing 200 g/kg DDGS; II– not supplemented, III – supplemented with feed enzymes (xylanase + phytase), IV – supplemented with sodium butyrate, V – supplemented with probiotic bacteria, VI – supplemented with herb extracts mixture, VII – supplemented with inulin, VIII – supplemented with chitosan.

Table 5. Effect of maize DDGS and experimental additives on concentrations of main groups of fatty acids in egg yolk lipids (%) Treatment I II III IV V VI VII VIII SEM

SFA 39.2 39.1 38.8 39.5 38.0 39.0 39.2 39.7 0.143

UFA 60.2 60.7 60.8 60.5 62.0 61.0 60.8 60.3 0.141

MUFA b

47.2 41.9a 41.4a 41.3a 42.8a 42.4a 42.2a 41.9a 0.273

PUFA a

13.1 18.8b 19.4b 19.1b 19.2b 18.6b 18.6b 18.4b 0.301

PUFA n-6 a

11.3 16.8b 17.5b 17.4b 17.4b 16.9b 16.9b 16.9b 0.298

PUFA n-3

n-6/n-3

1.78 1.84 1.85 1.87 1.88 1.85 1.83 1.84 0.024

6.36a 9.13b 9.50b 9.30b 9.26b 9.14b 9.23b 9.18b 0.179

a,b Within a column, values not sharing a common superscript letter are significantly different (P ≤ 0.05). Treatments: I – control diet, II–VIII – experimental diets containing 200 g/kg DDGS; II – not supplemented, III – supplemented with feed enzymes (xylanase + phytase), IV – supplemented with sodium butyrate, V – supplemented with probiotic bacteria, VI – supplemented with herb extracts mixture, VII – supplemented with inulin, VIII – supplemented with chitosan.

conversion (Świątkiewicz and Koreleski, 2006). In another study, Lumpkins et al. (2005) found no significant differences in the majority of egg

production and egg quality parameters between hens given a diet with 0 or 150 g/kg of DDGS obtained from modern ethanol production.

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Incorporation of 150 g/kg DDGS into the low energy diet decreased egg production from 26 to 34 weeks of age, but there was no negative effect of DDGS on performance after 34 weeks of age. Based upon these results the authors concluded that DDGS is an acceptable feed ingredient for laying hens and it could be used in layers’ commercial diets at a 100 to 120 g/kg inclusion rate. They suggested, however, a lower inclusion rate in diets with decreased energy content. There were no effects of a high dietary concentration of DDGS on most of the egg quality parameters. However, in hens given diets with 200 g/kg of DDGS the yolk colour score was significantly increased. This positive effect is probably connected with the high content of carotenoids found in DDGS, the concentration of which could have increased during the fermentation process. A positive effect of maize DDGS on egg yolk colour was also observed by Roberson et al. (2005), who found that yolk colour score increased quickly with a 100 g/kg DDGS diet. It is also consistent with other studies (Świątkiewicz and Koreleski, 2006; Loar et al., 2010; Shalash et al., 2010; Masa’deh et al., 2011; Krawczyk et al., 2012). In the present study, the dietary inclusion of 200 g/kg DDGS caused no changes in eggshell quality parameters. This is agreement with the results of previous studies conducted with layers (Lumpkins et al., 2005; Roberson et al., 2005; Świątkiewicz and Koreleski, 2006; Koreleski et al., 2011). The changes in yolk lipid profile observed in hens given a high DDGS diet were rather unfavourable for the dietetic value of eggs, because of an increase of cholesterol content and PUFA n-6/ PUFA n-3 ratio. Similar results were reported by Krawczyk et al. (2012), who found that the yolks of eggs from layers given the DDGS diet had increased n-6/n-3 ratio as compared to the control groups. This increase was accompanied by an increasing content of oleic and linoleic acids. In a study of Cheon et al. (2008) a 200 g/kg dietary concentration of DDGS decreased the content of oleic acid and increased the content of linoleic acid in the yolk, but had no effect on the concentration of unsaturated fatty acids. Heincinger et al. (2012), in an experiment with growing turkeys, reported that the breast meat lipids of birds given diets with 150 g/kg DDGS were characterised by a numerically higher PUFA n-6/PUFA n-3 ratio and contained more polyunsaturated fatty acids as compared to the control group. The effect of selected feed additives in the diets with high DDGS inclusion Despite the fact that 200 g/kg dietary concentration of DDGS had no negative influence on laying performance in the present study, positive effects of dietary enzymes, inulin and chitosan in diets

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with high DDGS on some egg production parameters were found. In our previous work the use of NSP-hydrolysing enzymes was effective in alleviating the negative effects of a high DDGS diet on laying performance and nutrient utilisation (Świątkiewicz and Koreleski, 2006, 2007). A positive effect of a DDGS rich diet supplemented with a NSP-hydrolysing enzyme preparation on egg performance parameters was found by Shalash et al. (2010) and Ghazalah et al. (2011). Similarly, the results of an experiment with broiler chickens suggested that adding carbohydrase to the diet containing a high concentration of DDGS can improve growth performance and digestibility of diet components in broilers (Liu et al., 2011). Olukosi et al. (2010) found that phytase improved nutrient utilisation in broilers given a DDGS-containing diet, but the combination of the carbohydrase and phytase enzymes did not produce a greater benefit than the use of phytase alone. In contrast, in another study with broilers Min et al. (2011) found no effect of adding a commercial enzyme preparation (carbohydrase activity) to a DDGS rich (300 g/kg) diet on weight gain, feed intake, feed conversion ratio and nutrient utilisation. The authors speculated that this lack of influence could be explained by the relatively old age of the broilers used in the experiment, while it is a well-known fact that exogenous enzyme effect is more pronounced during the early phase of the rearing period. To date there are no available results in the scientific literature on the effects of addition of inulin or chitosan to a diet containing a high concentration of DDGS. Because of its β-glycosidic bonds, inulin is not digested by the enzymes of monogastric animals, so it is completely available for fermentation by the intestinal microflora and may selectively stimulate the growth of lactic acid bacteria, improving the overall health status and performance in poultry. Corresponding with our findings were the results reported by Chen et al. (2005b) in layers given a maize-soybean diet. They found that incorporation of fructans with prebiotic properties to the diet significantly improved egg production and feed conversion ratio. In contrast, Yildiz et al. (2006) and Świątkiewicz et al. (2010) found no effect of inulin, added to a cereal-soybean meal diet, on laying performance parameters. Chitosan is a water soluble N-deacetylated product of chitin, which is well digested by birds (Hirano et al., 1990). The beneficial effect of chitosan on laying performance, found in our study, may be attributed to its influence on intestinal morphology and the presence of hypertrophied villi and epithelial cells in birds given a diet supplemented with chitosan (Khambualai et al., 2009). Similar, positive effects of chitosan on performance indices and N retention were observed in experiments with broiler chickens and ducks (Shi et al., 2005; Yuan and Chen, 2012).

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As observed in this study, the positive effect of inulin on some eggshell quality parameters is consistent with our previous work (Świątkiewicz et al., 2010) where prebiotic fructans beneficially affected eggshell quality parameters (eggshell percentage, density and breaking strength) in older hens (at 46, 58 and 70 weeks of age). Also, the results obtained by Chen and Chen (2004) were similar to our findings; they reported that supplementing the diet with 10 g/kg inulin increased eggshell percentage and eggshell breaking strength. This positive effect on eggshell quality is probably connected with the stimulation of mineral availability. Ortiz et al. (2009) reported that a diet supplementation with inulin increased retention of Ca, Zn and Cu. According to ScholzAhrens et al. (2007) the mechanism of the positive effect of fructans on mineral utilisation is complex and can be attributed to factors such as: the high solubility of minerals because of the increased production of short chain fatty acids by probiotic bacteria, an alteration of intestinal mucosa and an increase of the absorption surface by means of the beneficial effect of bacterial fermentation products on the proliferation of enterocytes, increased expression of Ca-binding proteins, release of bone modulating factors, a degradation of phytates by probiotic bacteria enzymes and an overall improvement of gut health. In this study, some of the used additives, i.e. herb extracts and, to a lesser extent, inulin and chitosan, decreased the cholesterol content in yolk lipids of hens given a diet with 200 g/kg DDGS. Such an influence of active substances contained in herbs leading to beneficial changes in the metabolism of lipids in the organism of laying hens and to a decrease of cholesterol concentration in blood and egg yolks, was previously found by, amongst others, Bolukbasi et al. (2010), Akbarian et al. (2011) and Islam et al. (2011). Similarly to our observations, Chen et al. (2005a) and Shang et al. (2010) found that prebiotic fructans, when added to a layers diet, decreased cholesterol concentration in yolk lipids. A similar effect was also reported when the diet was supplemented with chitosan (Nogueira et al., 2003). Likewise, a beneficial influence of chitosan on lipid metabolism was found in broiler chickens and this was connected with a decrease of cholesterol and triglycerides in the blood of the birds (Razdan and Pettersson, 1994; Razdan et al., 1997). In conclusion, the results of this study indicate that DDGS may be incorporated to a concentration of 200 g/kg in a well-balanced diet for laying hens without any detrimental effects on egg performance and egg quality, and such additives as feed enzymes (xylanase + phytase), inulin and chitosan can positively affect the performance of layers given the diets with a high concentration of DDGS.

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