<table><caption>**Table 2** Apparent ileal digestibility of crude protein, starch, amino acids and AME<sub>N</sub> level in five faba bean cultivars in raw and extruded forms for broilers<sup>1</sup></caption><thead><tr><th>Cultivars</th><th>Extrusion</th><th>AME<sub>N</sub><br>MJ/kg</th><th>CP<br>%</th><th>Lys<br>%</th><th>Thr<br>%</th><th>Starch<br>%</th></tr></thead><tbody><tr><td></td><td>–</td><td>10.83<sup>b</sup></td><td>85.9<sup>b</sup></td><td>89.7<sup>b</sup></td><td>81.6<sup>b</sup></td><td>77.2<sup>b</sup></td></tr><tr><td></td><td>+</td><td>14.97<sup>a</sup></td><td>89.5<sup>a</sup></td><td>94.7<sup>a</sup></td><td>85.0<sup>a</sup></td><td>96.9<sup>a</sup></td></tr><tr><td>Merlin</td><td></td><td>13.96<sup>a</sup></td><td>87.7<sup>a</sup></td><td>92.0<sup>a</sup></td><td>83.9<sup>a</sup></td><td>86.8<sup>bc</sup></td></tr><tr><td>Olga</td><td></td><td>12.86<sup>b</sup></td><td>87.5<sup>a</sup></td><td>92.4<sup>a</sup></td><td>84.2<sup>a</sup></td><td>87.0<sup>b</sup></td></tr><tr><td>Albus</td><td></td><td>11.00<sup>c</sup></td><td>84.0<sup>b</sup></td><td>89.8<sup>b</sup></td><td>77.2<sup>b</sup></td><td>84.0<sup>c</sup></td></tr><tr><td>Amulet</td><td></td><td>13.21<sup>b</sup></td><td>89.4<sup>a</sup></td><td>92.5<sup>a</sup></td><td>85.7<sup>a</sup></td><td>90.1<sup>a</sup></td></tr><tr><td>Kasztelan</td><td></td><td>13.81<sup>a</sup></td><td>90.0<sup>a</sup></td><td>94.2<sup>a</sup></td><td>85.4<sup>a</sup></td><td>86.4<sup>bc</sup></td></tr><tr><td><strong>SEM</strong></td><td></td><td>0.26</td><td>0.58</td><td>0.55</td><td>0.89</td><td>1.52</td></tr><tr><td><strong>Cultivar</strong></td><td></td><td>&lt; 0.001</td><td>&lt; 0.001</td><td>0.002</td><td>0.002</td><td>0.025</td></tr><tr><td><strong>Extrusion</strong></td><td></td><td>&lt; 0.001</td><td>&lt; 0.001</td><td>&lt; 0.001</td><td>0.018</td><td>&lt; 0.001</td></tr><tr><td><strong>Cultivar × Extrusion</strong></td><td></td><td>&lt; 0.001</td><td>0.160</td><td>0.004</td><td>0.028</td><td>&lt; 0.001</td></tr></tbody></table>

¹n = 6

<table><caption>**Table 3** Pearson's correlation coefficients between phytic P and resistant starch content in faba bean seed traits in raw and extruded forms and determined parameters (AID of starch and amino acids; n = 60, AME<sub>N</sub>; n = 100)</caption><thead><tr><th>Faba bean traits</th><th>Response</th><th>Correlation coefficient (r)</th><th>p</th></tr></thead><tbody><tr><td rowspan="4">Phytic – P content</td><td>AME<sub>N</sub></td><td>-0.89</td><td>&lt; 0.001</td></tr><tr><td>Starch AID</td><td>-0.88</td><td>&lt; 0.001</td></tr><tr><td>Lys AID</td><td>-0.70</td><td>&lt; 0.001</td></tr><tr><td>Thr AID</td><td>-0.46</td><td>&lt; 0.001</td></tr><tr><td rowspan="2">Resistant starch</td><td>AME<sub>N</sub></td><td>-0.80</td><td>&lt; 0.001</td></tr><tr><td>Starch AID</td><td>-0.93</td><td>&lt; 0.001</td></tr></tbody></table>

## Conclusions

Extrusion cooking is one of the processes which can improve the nutritional value of faba beans through a decrease in phytic acid, trypsin inhibitors and resistant starch content. Effect of extrusion cooking depend on the concentration of resistant starch and phytic P content in faba bean seeds.

## References

ALONSO, R.; AGUIRRE, A.; MARZO, F., 2000: Effects of extrusion and traditional processing methods on antinutrients and in vitro digestibility of protein and starch in faba and kidney beans. Food Chem. **68**, 159–165.

DIAZ, D.; MORLACCHINI, M.; MASOERO, F.; MOSCHINI, M.; FUSCONI, G.; PIVA, G., 2006: Pea seeds (Pisum sativum), faba beans (Vicia faba var. minor) and lupin seeds (Lupinus albus var. multitalia) as protein sources in broiler diets: effect of extrusion on growth performance. Ital J Anim. Sci. **5**, 43–53.

WOYENGO, T. A.; NYACHOTI, C. M., 2012: Ileal digestibility of amino acids for zero-tannin faba bean (Vicia faba L.) fed to broiler chicks. Poultry Sci. **91**, 439–443.
P-146 (ID 174)

EFFECT OF DIETARY SUPPLEMENTATION WITH RICE BRAN EXTRACTS ON THE GROWTH PERFORMANCE,
BLOOD PARAMETERS OF BROILER CHICKENS

H. K. Kang, J.H. Kim, C. H. Kim, I. S. Jeon, J. Hwangbo

Poultry Science Division, Livestock Resource Development, National Institute of Animal Science, RDA, Seonghwan
330-801, Korea

**Abstract**

The objective of this experiment was to investigate the effect of rice bran extracts (RBE) as the dietary supplementation on growth performance, blood parameter, and immune response in broiler chickens. Two hundred forty 1-day-old ROSS 308 male broilers were allotted to 4 dietary treatments with 6 replicated pens consisting of 10 chicks. Three additional diets were prepared by adding 5, 10 or 20 g/kg of RBE to the basal diet. The experimental diets were fed on an *ad libitum* basis to the birds during 35 d. Consequently, increase in the inclusion level of RBE in diets improved BW gain by 9.0, 7.8, or 7.3%, respectively. Feed conversion ratio of birds by 10.3, 11.4, or 14.3%, respectively, but affected neither the feed intake of birds nor erythrocytes of birds. However, heterophil, lymphocyte, and monocytes increased (linear and quadratic, P<0.01) with inclusion level of RBE in diets increased. As expected, increasing inclusion level of RBE in diets increased (linear, P<0.01) the concentrations of total cholesterol. AST, ALT, glucose, and albumin were not affected by inclusion of RBE in diets. Increasing inclusion level of RBE in diets increased concentrations of IgG (linear, P<0.01). There was no effect of inclusion level of RBE in diets on concentrations of IgM. These results suggest that dietary RBE may be used as a functional ingredient to improve growth performance, total cholesterol, and immune response of birds.

**Keywords:** broiler, rice bran extracts, growth performance, total cholesterol

**Introduction**

Rice is the principle cereal food in Korea, and the by-products from its milling are used as important feed resources. One such by-product is rice bran extracts (RBE); it is gaining commercial importance worldwide because of its many beneficial nutritive and biological effects. Rice bran oil contains 90-96% saponifiable lipids and about 4% unsaponifiable lipids. The saponifiable lipids include 68-71% triglycerides, 2-3% diglycerides, 5-6% monoglycerides, 2-3% free fatty acids (FFAs), 2-3% waxes, 5-7% glycolipids, and 3-4% phospholipids (McCaskill and Zhang, 1999), whereas the principle component of the unsaponifiable fraction is *r*-oryzanol (Raghuram and Rukmini, 1991).

*r*-oryzanol, a phytosteryl ferulate mixture extracted from rice bran oil, has a wide spectrum of biological activities, as well as antioxidant properties (Juliano et al., 2005). RBE has an excellent fatty acid profile. Three major fatty acids, palmitic, oleic, and linoleic account for 90% of the total fatty acids of rice bran oil (Amarasinghe and Gangodavilage, 2004). The crude form of rice bran extracts is known to improve the growth performance of broilers. Dietary supplementation of rice bran oil in broilers improved growth performance (Anitha et al., 2006; Purushothaman et al., 2005) and decreased cholesterol concentrations (Berger et al., 2005). However, data pertaining to the effects of dietary supplementation of RBE on the growth performance, blood parameters, and immune response of animals are limited. In this regard, we investigated the effect of dietary supplementation of RBE on the growth performance, blood parameters, and immune response of broilers.

**Material and Methods**

A total of two hundred forty 1-day-old ROSS 308 (initial BW = 36 ± 0.74 g) were obtained from local hatchery (Samhwa Hatchery, Hongseong, Republic of Korea) and were housed in battery cages (86 cm × 57 cm × 35 cm, width × length × height) in an environmentally controlled room. The experiment was performed as a completely randomized design with 6 replicate pens consisting of 10 chicks. A 2-phase feeding program with a starter diet from 0 to 21 d and a grower diet from 22 to 35 d was used in this experiment. Within each phase, a control diet was formulated to meet or exceed NRC (1994) requirements of broiler chickens for macro- and micronutrients. Three additional diets were prepared by adding 5, 10 or 20 g/kg of RBE to the basal diet. The RBE was added at the expense of the basal diet. The experimental diets were mash form. The diets and water were available *ad libitum*. The room temperature for both conventional and battery cages was maintained at 30°C during the first week of the experiment and then gradually decreased to
24°C at the end of the experiment as recommended by the Ross manual. A 24-h lighting schedule was used throughout the experiment. All data were analyzed by ANOVA according to completely randomized design (Steel et al., 1997) using the PROC MIXED procedure of SAS (SAS Institute Inc., Cary, NC). Outlier data were identified according to Steel et al. (1997), using the UNIVARIATE procedure of SAS, but no outliers were detected. The battery cage was an experimental unit for growth performance data, whereas the individual bird was an experimental unit for blood parameter, serum enzyme activities and immune response data. Dietary treatment was fixed effect in all statistical models. The LSMEANS procedure was used to calculate mean values. The orthogonal polynomial contrast test was performed to determine linear and quadratic effects of increasing inclusion level of RBE in diets on each measurement. Significance and tendency for statistical tests were set at P < 0.05 and 0.05 ≤ P ≤ 0.10, respectively.

## Results

During the initial 21 d of the experiment, BW gain and feed conversion ratio increased (linear, P<0.05) with inclusion level of RBE, but feed intake was not influenced by inclusion of RBE in diets. During 22 to 35 d of the experiment, BW gain increased (linear and quadratic, P<0.05) with inclusion level of RBE and a tendency for improved feed conversion ratio (linear, P<0.10) was observed as inclusion level of RBE in diets increased. There was no effect of inclusion level of RBE in diets on feed intake of birds. For overall experiment, increasing inclusion level of RBE in diets increased BW gain (linear and quadratic, P<0.01), and improve feed conversion ratio (linear, P<0.05). There was no effect of inclusion level of RBE in diets on feed intake of birds. There was no effect of inclusion level of RBE in diets on erythrocytes of birds. However, heterophil, lymphocyte, and monocytes increased (linear and quadratic, P<0.01) with inclusion level of RBE in diets increased (Table 4). As expected, increasing inclusion level of RBE in diets increased (linear, P<0.01) the concentrations of total cholesterol (Table 5). AST, ALT, glucose, and albumin were not affected by inclusion of RBE in diets. Increasing inclusion level of RBE in diets increased concentrations of IgG (linear, P<0.01). There was no effect of inclusion level of RBE in diets on concentrations of IgM.

<table><caption>**Table 1.** Immune response of broiler chickens fed the diet containing rice bran extract (RBE)ª</caption><thead><tr><th rowspan="2">Items</th><th colspan="4">RBE (g/kg)</th><th rowspan="2">Pooled SEM</th><th colspan="2">P-value</th></tr><tr><th>0</th><th>5</th><th>10</th><th>20</th><th>Linear</th><th>Quadratic</th></tr></thead><tbody><tr><td>IgG (mg/mL)</td><td>1.26</td><td>1.56</td><td>1.56</td><td>2.51</td><td>0.51</td><td>0.01</td><td>0.27</td></tr><tr><td>IgM (µg/mL)</td><td>132.2</td><td>259.7</td><td>162.6</td><td>225.9</td><td>82.18</td><td>0.63</td><td>0.71</td></tr></tbody></table>

## Conclusions

The results of this study indicate that dietary supplementation of rice bran extracts improves growth performance and decreases total cholesterol level in broilers. Therefore, dietary rice bran extracts is considered a valuable functional ingredient to improve the growth performance of birds.

## Literature

ANITHA, B., MOORTHY, M., VISWANATHAN, K., 2006. Production performance of broilers fed with crude rice bran oil. Int. J. Poult. Sci., **5**: 1046-1052.

BACH KNUDSEN, K. E. 2001. Development of antibiotic resistance and options to replace antimicrobials in animal diets. Proc. Nutr. Soc. **60**: 291-299.

CAO, F. L., ZHANG, X. H., ZHAO L. G., T. WANG. 2012. Effect of feeding fermented Ginko biloba leaves on growth performance, meat quality, and lipid metabolism in broilers. Poult. Sci. **91**: 1210-1221.

DINI, H. 2010. The nutrient evaluation of fermented rice bran as poultry feed. Int. J. Poult. Sci. **9**: 152-154.

JULIANO, C., COSSU, M., ALAMANNI M. C., PIU, L. 2005. Antioxidant activity of gamma oryzanol: mechanism of action and its effect on oxidative stability of pharmaceutical oils. Int. J. Pharm. **299**: 146-154.

RUKMINI, C., RAGHURAM, T. C. 1991. Nutritional and biochemical aspects of the hypolipidemic action of rice bran oil: a review. J. Am. Coll. Nutr. **10**: 593-601.

<footnote>ªData are least squares means of 6 observations per treatments.</footnote>
P-148 (ID 127)

USE OF SEMI-LEAFLESS PEAS (PISUM SATIVUM L) WITH ENZYME ADDITION IN DIETS FOR BROILERS

E. Koivunen¹*, P. Tuunainen¹, E. Talvio², E. Valkonen³, T. Tupasela⁴, J. Valaja⁵

¹Pig and Poultry Production, Green Technology, Natural Resources Institute Finland (Luke), Jokioinen, Finland

²HKScan Finland Oy, Turku, Finland

³Hankkija Oy, Hyvinkää, Finland

⁴New Products, Services and Technology, New Business Opportunities, Natural Resources Institute Finland (Luke), Jokioinen, Finland

⁵Dept. of Agricultural Sciences, University of Helsinki, Helsinki, Finland

*Corresponding author: erja.koivunen@luke.fi

**Abstract**

The trial was conducted to study the effects of dietary pea inclusion and the supplementation of combination of xylanase (XLS), amylase (AMS) and protease (PRT) on broiler performance, intestinal viscosity and organoleptic quality of meat. A total of 3000 1-d-old Ross 508 broiler chickens were reared. Birds were sexed and equal number of females and males were reared in each experimental unit. Eight experimental diets were arranged 4×2 factorially, the factors being dietary pea inclusion (0, 150, 300 and 450 g/kg) and the supplementation of combination of XLS, AMS and PRT (0.1 % in the diet). Pea inclusion had no effect (*P* > 0.05) on body weight, body weight gain and feed consumption. FCR of the birds fed XLS, AMS and PRT supplemented diets increased (*P* ≤ 0.042) along pea inclusion. Pea inclusion improved (*P* ≤ 0.038) the tenderness and juiciness of breast meat for the XLS, AMS and PRT supplemented diets. The use of XLS, AMS and PRT improved the nutrition value of pea diets as evidenced from the improvement in body weight especially for the birds fed control diet (*P* < 0.001) and diet included peas up to 150 g/kg (*P* = 0.005). The supplementation of XLS, AMS and PRT improved FCR and decreased (*P* = 0.040) feed consumption of the birds fed control diet (*P* = 0.044). These results suggest that XLS, AMS and PRT improved more the nutritional value of wheat than that of pea. In conclusion, 450 g/kg semi-leafless peas studied can be used in the broiler grower diets without negative effects on the bird performance. The use of combination of XLS, AMS and PRT improve bird performance.

**Keywords:** amylase, broiler, pea, protease, xylanase

**Introduction**

There is a strong interest in maximizing the use of locally produced vegetable protein sources, like peas (*Pisum sativum* L.), as a substitute for imported SBM. The different combinations of enzymes have been studied to improve nutritive value of peas (BRENES et al., 1993; IGBASAN & GUENTER 1996; COWIESON et al., 2003). However, there is a lack of studies where benefits of the addition of the combination xylanase (XLS), amylase (AMS) and protease (PRT) to wheat-pea based diets would have been evaluated. The aim of this study was to study the effects of dietary pea inclusion and the supplementation of combination of XLS, AMS and PRT on broiler performance, intestinal viscosity and organoleptic quality of the meat. A further aim of this study was to find an optimal inclusion level of peas (cv. Karita) in diets for broilers.

**Materials and Methods**

A total of 3000 1-d-old Ross 508 broiler chickens were reared in 48 floor pens with equal number of females and males. Eight wheat-SBM based experimental diets were arranged 4×2 factorially, the factors being dietary pea (cv. Karita) inclusion (0, 150, 300 and 450 g/kg) and the supplementation of combination of XLS, AMS and PRT (0.1 % in the diet). The exogenous enzyme combination used was Avizyme 1200 -enzyme (Danisco Animal Nutrition, Marlborough, UK). After slaughtering one breast meat of one bird per replicate was removed and frozen for organoleptic quality evaluation. The ileal digesta of one bird per replicate was collected from the distal part of ileum for viscosity determination. The experimental data was subjected to ANOVA using the GLM procedure of SAS (SAS Institute Inc., Cary, NC, USA). The treatment effects were separated into six orthogonal contrasts. In the current study, *P* ≤ 0.05 was considered to be significant.
## Results and Discussion

Dietary pea inclusion up to 450 g/kg had no effect (P > 0.05) on body weight, carcass weight, body weight gain or feed consumption. Bird performance was comparable to that presented in earlier studies, where pea inclusion ranged from 100 to 480 g/kg (IGBASAN & GUENTER 1996; LAUDADIO & TUFARELLI 2010; DOTAS et al., 2014). Pea inclusion impaired (P ≤ 0.042) the FCR of birds on XLS, AMS and PRT supplemented diets in line with the results by COWIESON et al., (2003) (Table 1). The addition of XLS, AMS and PRT increased final body weight (P < 0.041) of the birds on control diet and diets with 150 g/kg and 450 g/kg of peas. Body weight gain (P < 0.040) of the birds on control diet and diets with 150 g/kg and 450 g/kg peas and FCR of the birds on control diet and diets with 150 and 300 g/kg peas improved (P ≤ 0.040) by the supplementation of XLS, AMS and PRT. The addition of XLS, AMS and PRT only decreased (P = 0.044) control diet consumption. The beneficial effects of XLS, AMS and PRT combination on bird performance were most likely a result of the effects of the enzymes on backbone substituent (arabinoxylans) in wheat. The intestinal viscosity of the birds (age 20 day) decreased (P ≤ 0.021) by the addition of XLS, AMS and PRT and for birds on unsupplemented diets along pea inclusion. Our results in the unchanged flavour of breast meat agree with the results of MC NEILL et al., (2004). Pea inclusion improved (P < 0.038) tenderness and juiciness of breast meat of the birds on XLS, AMS and PRT supplemented diets.

## Conclusions

When diets are balanced with regards to their amino acid content 450 g/kg semi-leafless peas can be used in broiler grower diets based on wheat and SBM. The use of enzyme combination of XLS, AMS and PRT enhanced the nutritive value of wheat-pea diets. The beneficial effects of this were however more likely a result of the effects of the enzymes on backbone substituent in wheat. The dietary pea inclusion improves the tenderness and juiciness of breast meat, but only for the XLS, AMS and PRT supplemented diets.

## References

* BRENES, A., ROTTER, B.A., MARQUARDT, R.R., GUENTER, W. 1993: The nutritional value of raw, autoclaved and dehulled peas (Pisum Sativum L.) in chicken diets as affected by enzyme supplementation. Can. J. Anim. Sci. **73**, 605–614.

* COWIESON, A.J., ACAMOVIC, T., BEDFORD, M.R. 2003: Supplementation of diets containing pea meal with exogenous enzymes: Effects on weight gain, feed conversion, nutrient digestibility and gross morphology of the gastrointestinal tract of growing broiler chicks. Brit. Poultry Sci. **44**, 427–437.

* IGBASAN, F.A., GUENTER, W. 1996: The evaluation and enhancement of the nutritive value of yellow-, green and brown-seeded pea cultivars for unpelleted diets given to broiler chickens. Anim Feed Sci. Tech. **63**, 9–24.

* LAUDADIO, V., TUFARELLI, V. 2010: Growth performance and carcass and meat quality of broiler chickens fed diets containing micronized-dehulled peas (Pisum Sativum cv. Spirale) as a substitute of soybean meal. Poult. Sci. **89**, 1537–1543.
<table><caption>**Table 1.** The effects of dietary pea inclusion level and enzyme supplementation on bird performance, ileal viscosity and the organoleptic quality of breast meat.</caption><thead><tr><th>Treatment</th><th>1</th><th>2</th><th>3</th><th>4</th><th>5</th><th>6</th><th>7</th><th>8</th><th>SEM</th><th>P<sub>I</sub></th><th>P<sub>I</sub></th><th>E<sub>control</sub></th><th>E<sub>15</sub></th><th>E<sub>30</sub></th><th>E<sub>45</sub></th></tr></thead><tbody><tr><td>Pea inclusion,<br>A<sup>AV</sup> enzyme 1200</td><td>0</td><td>150</td><td>300</td><td>450</td><td>0</td><td>150</td><td>300</td><td>450</td><td></td><td>-</td><td>+</td><td></td><td></td><td></td><td></td></tr><tr><td>Body weight, g</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>d 9</td><td>238</td><td>239</td><td>240</td><td>238</td><td>238</td><td>243</td><td>239</td><td>244</td><td>3.0</td><td>0.858</td><td>0.406</td><td>0.951</td><td>0.254</td><td>0.842</td><td>0.183</td></tr><tr><td>d 37</td><td>2263</td><td>2288</td><td>2267</td><td>2265</td><td>2313</td><td>2352</td><td>2300</td><td>2305</td><td>13.6</td><td>0.684</td><td>0.130</td><td>0.014</td><td>0.002</td><td>0.097</td><td>0.041</td></tr><tr><td>Body weight gain, g</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>from d 1 to d 9</td><td>198</td><td>198</td><td>200</td><td>197</td><td>197</td><td>203</td><td>199</td><td>203</td><td>2.9</td><td>0.946</td><td>0.368</td><td>0.898</td><td>0.297</td><td>0.879</td><td>0.164</td></tr><tr><td>from d 10 to d 37</td><td>2025</td><td>2049</td><td>2027</td><td>2026</td><td>2075</td><td>2108</td><td>2060</td><td>2061</td><td>13.8</td><td>0.659</td><td>0.095</td><td>0.015</td><td>0.004</td><td>0.093</td><td>0.080</td></tr><tr><td>from d 1 to d 37</td><td>2223</td><td>2247</td><td>2226</td><td>2224</td><td>2272</td><td>2311</td><td>2259</td><td>2265</td><td>13.6</td><td>0.665</td><td>0.131</td><td>0.014</td><td>0.002</td><td>0.094</td><td>0.040</td></tr><tr><td>Feed consumption,<br>from d 1 to d 9</td><td>25</td><td>25</td><td>25</td><td>25</td><td>25</td><td>25</td><td>25</td><td>25</td><td>0.3</td><td>0.677</td><td>0.340</td><td>0.116</td><td>0.847</td><td>0.468</td><td>0.833</td></tr><tr><td>from d 10 to d 37</td><td>131</td><td>129</td><td>129</td><td>130</td><td>128</td><td>130</td><td>128</td><td>131</td><td>1.0</td><td>0.434</td><td>0.370</td><td>0.057</td><td>0.458</td><td>0.295</td><td>0.405</td></tr><tr><td>from d 1 to d 37</td><td>104</td><td>103</td><td>103</td><td>103</td><td>102</td><td>103</td><td>102</td><td>104</td><td>0.8</td><td>0.516</td><td>0.305</td><td>0.044</td><td>0.783</td><td>0.288</td><td>0.625</td></tr><tr><td>FCR, g of feed/g gr</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>from d 1 to d 9</td><td>1.15</td><td>1.14</td><td>1.14</td><td>1.13</td><td>1.13</td><td>1.12</td><td>1.13</td><td>1.12</td><td>0.013</td><td>0.427</td><td>0.831</td><td>0.205</td><td>0.260</td><td>0.657</td><td>0.334</td></tr><tr><td>from d 10 to d 37</td><td>1.85</td><td>1.79</td><td>1.82</td><td>1.83</td><td>1.77</td><td>1.75</td><td>1.77</td><td>1.82</td><td>0.011</td><td>0.567</td><td>0.002</td><td>&lt;0.001</td><td>0.009</td><td>0.003</td><td>0.436</td></tr><tr><td>from d 1 to d 37</td><td>1.79</td><td>1.73</td><td>1.76</td><td>1.77</td><td>1.71</td><td>1.69</td><td>1.71</td><td>1.75</td><td>0.009</td><td>0.490</td><td>0.003</td><td>&lt;0.001</td><td>0.005</td><td>0.003</td><td>0.309</td></tr><tr><td>Ileal viscosity<sup>2</sup>, cPs</td><td>10.2</td><td>7.3</td><td>7.2</td><td>6.8</td><td>3.5</td><td>3.9</td><td>3.5</td><td>3.3</td><td>0.90</td><td>0.021</td><td>0.749</td><td>&lt;0.001</td><td>0.011</td><td>0.006</td><td>0.008</td></tr><tr><td>Ileal viscosity<sup>3</sup>, cPs</td><td>6.6</td><td>6.6</td><td>5.9</td><td>5.7</td><td>5.0</td><td>4.5</td><td>4.4</td><td>4.5</td><td>0.47</td><td>0.090</td><td>0.461</td><td>0.017</td><td>0.004</td><td>0.036</td><td>0.073</td></tr><tr><td>Tenderness</td><td>4.8</td><td>4.8</td><td>4.8</td><td>4.8</td><td>4.1</td><td>4.9</td><td>4.5</td><td>5.3</td><td>0.25</td><td>0.932</td><td>0.016</td><td>0.051</td><td>0.635</td><td>0.344</td><td>0.159</td></tr><tr><td>Juiciness</td><td>4.5</td><td>4.5</td><td>4.5</td><td>4.7</td><td>4.2</td><td>4.6</td><td>4.5</td><td>4.8</td><td>0.17</td><td>0.532</td><td>0.038</td><td>0.166</td><td>0.889</td><td>0.889</td><td>0.779</td></tr><tr><td>Flavour</td><td>4.6</td><td>4.8</td><td>4.7</td><td>4.7</td><td>4.3</td><td>4.9</td><td>4.4</td><td>4.7</td><td>0.19</td><td>1.000</td><td>0.614</td><td>0.262</td><td>0.802</td><td>0.262</td><td>0.900</td></tr></tbody></table>

P<sub>I</sub> = the linear effect of pea inclusion

E<sub>control</sub> = the effect enzyme supplementation on control diet (treatment 1 vs. treatment 5)

E<sub>15</sub> = the effect enzyme supplementation on diet containing peas 150 g/kg (treatment 2 vs. treatment 6)

E<sub>30</sub> = the effect enzyme supplementation on diet containing peas up to 300 g/kg (treatment 3 vs. treatment 7)

E<sub>45</sub> = the effect enzyme supplementation on diet containing peas up to 450 g/kg (treatment 4 vs. treatment 8)

<footnote n="1">¹entire experiment</footnote>

<footnote n="2">²at 20 d of age</footnote>

<footnote n="3">³at 30 d of age</footnote>
# NATURAL STABILIZATION OF RICE BRAN AND THE EFFECT ON BROILER PERFORMANCE AND CARCASS YIELD

A. A. da S. Catalan<fnref n="1" />, E. Gopinger<fnref n="1" />, V. S. de Avila<fnref n="2" />, D. Surek<fnref n="2" />, L. dos S. Lopes<fnref n="2" />, E. L. Krabbe<fnref n="2" />

**Corresponding author:** everton.krabbe@embrapa.br

## Abstract

Rice bran represents eight percent of the raw rice grain processing, and contains a considerable nutrient profile for broiler. However, due to natural enzymes present at the external part of the rice grain and consequently concentrated in the bran portion when rice is submitted to polishing, results is a very unstable feedstuff, with fast rancidity generation during storage, subsequent oxidation and consequently negative effect on poultry performance. Alternatives for rice bran stabilization are of interest, especially natural alternatives. In this sense, 1000 ppm soybean tocopherols, 1000 ppm neem extract (Azadirachta indica) and 250 ppm citric acid were evaluated straight or in combination (T1 – Control, T2 - Neem; T3 – Citric Acid; T4 - Neem + Citric Acid; T5 – Tocopherols; T6 – Tocopherols + Neem; T7 – Tocopherols + Citric Acid; T8 – Tocopherols + Neem + Citric Acid), for rancidity or oxidation prevention, during 90 days storage period. Subsequently, those rice brans were tested in broiler diets to measure the effect of additives/stabilization programs on performance and carcass yield. Experimental diets contained 6.5% rice bran, fed from one to 28 d of age. Feed intake, weight gain and feed conversion ratio were not affected by the different rice bran stabilization programs. At 28 d, two birds per replicate were slaughtered and breast, leg and liver weight measured. In terms of yields, no difference between treatments was observed.

**Keywords:** Bird, Nutrition, alternative feedstuff, oxidation, feed efficiency

## Introduction

Rice bran is a by-product of rice milling and features nutrients such as oil, including essential fatty acids, protein, fiber, vitamins, and antioxidants, such as g-oryzanol. However, in Brazil this by-product has a low commercial value, enabling its use mainly as an ingredient in animal feed, but because of the highly unstable nature during storage, and high in lipids, results in its rancidity, thus requiring some care. The evaluation of natural antioxidants for rice bran stabilization is scarce, therefore, the aim of this study was to evaluate the effect of different natural antioxidant additives in stored rice bran and later incorporated into the chicken diet cut to evaluate the performance and carcass yield.

## Material and Methods

**Place of execution:** The research was performed at Embrapa Swine and Poultry, Concórdia, Santa Catarina State, Brazil. The rice bran (FAI) was purchased directly from the rice mill and immediately transferred to Embrapa R&D Feed Plant Facility for preservative program implementation.

**Rice bran treatments and storage:** To avoid the rancidity or oxidation during storage, FAI was treated with 1000 ppm soybean tocopherols, 1000 ppm neem extract (Azadirachta indica) and 250 ppm citric acid were evaluated straight or in combination, like demonstration on Table 1 (T1 – Control, T2 – Neem; T3 – Citric Acid; T4 - Neem + Citric Acid; T5 – Tocopherols; T6 – Tocopherols + Neem; T7 – Tocopherols + Citric Acid; T8 – Tocopherols + Neem + Citric Acid). All application were done in a typical feed mixer, were liquid additives were sprayed using a membrane pump and a spray nozzle. The mixing time after ending application was 10 minutes, as SOP description for this machine. All brans were stored in raffia bags of 15 kg. All treatments and replicates were kept under same condition, for 90 days. The climate conditions during the storage period were, maximum and minimum, temperature of 32 and 5Cº and relative humidity 95 to 35%, respectively, and an average rainfall of 215mm / month.

<footnote n="1">¹Dept. of Animal Science, Federal University of Pelotas, Pelotas-RS/Brazil</footnote>

<footnote n="2">²Brazilian Agricultural Research Corporation, Swine & Poultry, Concórdia-SC/Brazil</footnote>
<table><caption>**Table 1.** Description of the treatment for rancidity prevention applied to rice bran (FAI)</caption><thead><tr><th rowspan="2">Identification</th><th colspan="3">Natural additive sources (ppm)</th></tr><tr><th>Neem (N)</th><th>Citric acid (CA)</th><th>Tocopherols (T)</th></tr></thead><tbody><tr><td>FAI Control</td><td>0</td><td>0</td><td>0</td></tr><tr><td>FAI N</td><td>1000</td><td>0</td><td>0</td></tr><tr><td>FAI CA</td><td>0</td><td>250</td><td>0</td></tr><tr><td>FAI CA+N</td><td>1000</td><td>250</td><td>0</td></tr><tr><td>FAI T</td><td>0</td><td>0</td><td>1000</td></tr><tr><td>FAI T+N</td><td>1000</td><td>0</td><td>1000</td></tr><tr><td>FAI T+CA</td><td>0</td><td>250</td><td>1000</td></tr><tr><td>FAI T+CA+N</td><td>1000</td><td>250</td><td>1000</td></tr></tbody></table>

**Feed Production:** Diet was produced according Table 2. All feed were produced in mash form. The nutritional profile followed the Brazilian standards for broiler (Rostagno et al., 2011). The experimental FAI was included at a 6.5% level, as recommended by same author.

<table><caption>**Table 2.** Experimental diet composition fed from one to 28 days of age, for broiler male, Cobb 500, containing rice bran (FAI) stored for 90 days, under distinct preservation treatments using natural additives (Neem - N, citric acid - CA e tocopherols - T).</caption><thead><tr><th>Ingredient</th><th>T1</th><th>T2</th><th>T3</th><th>T4</th><th>T5</th><th>T6</th><th>T7</th><th>T8</th></tr></thead><tbody><tr><td>Corn (7.5%)</td><td>50.55</td><td>50.55</td><td>50.55</td><td>50.55</td><td>50.55</td><td>50.55</td><td>50.55</td><td>50.55</td></tr><tr><td>Soybean Meal (45%)</td><td>36.07</td><td>36.07</td><td>36.07</td><td>36.07</td><td>36.07</td><td>36.07</td><td>36.07</td><td>36.07</td></tr><tr><td>FAI Control</td><td>6.50</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><td>FAI N</td><td>0</td><td>6.50</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><td>FAI CA</td><td>0</td><td>0</td><td>6.50</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><td>FAI CA+N</td><td>0</td><td>0</td><td>0</td><td>6.50</td><td>0</td><td>0</td><td>0</td><td>0</td></tr><tr><td>FAI T</td><td>0</td><td>0</td><td>0</td><td>0</td><td>6.50</td><td>0</td><td>0</td><td>0</td></tr><tr><td>FAI T+N</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>6.50</td><td>0</td><td>0</td></tr><tr><td>FAI T+CA</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>6.50</td><td>0</td></tr><tr><td>FAI T+CA+N</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>0</td><td>6.50</td></tr><tr><td>Soybean oil</td><td>2.92</td><td>2.92</td><td>2.92</td><td>2.92</td><td>2.92</td><td>2.92</td><td>2.92</td><td>2.92</td></tr><tr><td>Kaolin (Filler)</td><td>0.10</td><td>0.10</td><td>0.10</td><td>0.10</td><td>0.10</td><td>0.10</td><td>0.10</td><td>0.10</td></tr><tr><td>Dical-Phosphate<sup>1</sup></td><td>1.38</td><td>1.38</td><td>1.38</td><td>1.38</td><td>1.38</td><td>1.38</td><td>1.38</td><td>1.38</td></tr><tr><td>Limestone<sup>2</sup></td><td>1.00</td><td>1.00</td><td>1.00</td><td>1.00</td><td>1.00</td><td>1.00</td><td>1.00</td><td>1.00</td></tr><tr><td>Common Salt</td><td>0.50</td><td>0.50</td><td>0.50</td><td>0.50</td><td>0.50</td><td>0.50</td><td>0.50</td><td>0.50</td></tr><tr><td>DL-methionine</td><td>0.27</td><td>0.27</td><td>0.27</td><td>0.27</td><td>0.27</td><td>0.27</td><td>0.27</td><td>0.27</td></tr><tr><td>L-lysine</td><td>0.22</td><td>0.22</td><td>0.22</td><td>0.22</td><td>0.22</td><td>0.22</td><td>0.22</td><td>0.22</td></tr><tr><td>Vitamin Premix<sup>3</sup></td><td>0.10</td><td>0.10</td><td>0.10</td><td>0.10</td><td>0.10</td><td>0.10</td><td>0.10</td><td>0.10</td></tr><tr><td>Mineral Premix<sup>4</sup></td><td>0.05</td><td>0.05</td><td>0.05</td><td>0.05</td><td>0.05</td><td>0.05</td><td>0.05</td><td>0.05</td></tr><tr><td>Toxin binder</td><td>0.18</td><td>0.18</td><td>0.18</td><td>0.18</td><td>0.18</td><td>0.18</td><td>0.18</td><td>0.18</td></tr><tr><td><strong>TOTAL</strong></td><td><strong>100</strong></td><td><strong>100</strong></td><td><strong>100</strong></td><td><strong>100</strong></td><td><strong>100</strong></td><td><strong>100</strong></td><td><strong>100</strong></td><td><strong>100</strong></td></tr><tr><td>AMEn (kcal/kg)</td><td>2950</td><td>2950</td><td>2950</td><td>2950</td><td>2950</td><td>2950</td><td>2950</td><td>2950</td></tr><tr><td>Crude Protein (%)</td><td>21.05</td><td>21.05</td><td>21.05</td><td>21.05</td><td>21.05</td><td>21.05</td><td>21.05</td><td>21.05</td></tr><tr><td>Calcium (%)</td><td>0.820</td><td>0.820</td><td>0.820</td><td>0.820</td><td>0.820</td><td>0.820</td><td>0.820</td><td>0.820</td></tr><tr><td>P available (%)</td><td>0.390</td><td>0.390</td><td>0.390</td><td>0.390</td><td>0.390</td><td>0.390</td><td>0.390</td><td>0.390</td></tr><tr><td>P total (%)</td><td>0.720</td><td>0.720</td><td>0.720</td><td>0.720</td><td>0.720</td><td>0.720</td><td>0.720</td><td>0.720</td></tr><tr><td>Sodium (%)</td><td>0.210</td><td>0.210</td><td>0.210</td><td>0.210</td><td>0.210</td><td>0.210</td><td>0.210</td><td>0.210</td></tr></tbody></table>

FAI – rice bran as presented at Table 1; ¹Calcium: minimum 210g/kg, maximum 250g/kg; Phosphorus: 180g/kg; ²Calcium: minimum 33%;

³Content per kg of product: Vitamin A: 9000000.000 UI, Vitamin D3: 2500000.00 UI, Vitamin E: 20000.00 UI, Vitamin K3: 2500.00mg, Vitamin B1: 1500.00mg, Vitamin B2: 6000.00mg, Vitamin B6: 3000.00mg, Vitamin B12: 12000.00mcg, Pantothenic Acid: 12g, Niacin: 25g, Folic Acid: 800.00mg, Biotin: 60.00mg, Selenium: 250.00mg. ⁴Specification per kg of product: Copper: 20g, Iron: 100g, Manganese: 160g, Cobalt: 2000.00mg, Iodine: 2000.00mg, Zinc: 100g.
**In vivo trial:** With the aim to evaluate the FAI nutritional quality after 90 d storage period under different preservation strategies, a broiler performance trial was performed, using 960 male Cobb500 birds in vertical metallic cages, equipped with a feeder and nipple drinkers, up to 28 days age. The birds were allocated in a randomized block design with eight treatments and 10 replicates of 12 birds each. Diets previously described were delivered ad libitum. At 7, 14, 21 and 28 days of age the feed intake, body weight, body weight gain, and feed conversion was determined. At 28 days, two birds, selected based on the mean body weight of the replicate, were selected, identified, and submitted to carcass part yield. All parameters were submitted to analysis of variation and means were compared throughout t-Student Test at a 5% of significance level.

## Results and conclusion

Feed intake, weight gain and feed conversion ratio were not affected by the different rice bran stabilization programs. In terms of carcass part yields, no difference between treatments was observed (Table 3).

<table><caption>**Table 3.** Broiler performance and carcass part yields, male, Cobb500, fed with diets containing rice bran (FAI) stored for 90 days and preserved with distinct natural additives (Neem – N, citric acid – CA and Tocopherols – T) (mean ± standard error).</caption><thead><tr><th>Treatments</th><th>FI (g)</th><th>WG (g)</th><th>FCR (g:g)</th><th>BW (g)</th><th>BY (%)</th><th>LiY (%)</th><th>LeY (%)</th></tr></thead><tbody><tr><td>FAI Control</td><td>2066±16.7</td><td>1434±22.0</td><td>1.450±0.010</td><td>1415±28.7</td><td>22.8± 0.32</td><td>2.100±0.055</td><td>12.8± 0.12</td></tr><tr><td>FAI N</td><td>2062±15.6</td><td>1439±10.4</td><td>1.434±0.007</td><td>1458±30.5</td><td>23.6± 0.29</td><td>1.987±0.046</td><td>13.0± 0.13</td></tr><tr><td>FAI CA</td><td>2047± 9.1</td><td>1426±19.9</td><td>1.436±0.016</td><td>1432±22.8</td><td>23.1± 0.41</td><td>1.977±0.031</td><td>13.0± 0.19</td></tr><tr><td>FAI CA+N</td><td>2077±18.9</td><td>1424± 8.6</td><td>1.460±0.015</td><td>1438±28.1</td><td>22.5± 0.29</td><td>1.965±0.039</td><td>13.1± 0.16</td></tr><tr><td>FAI T</td><td>2057±20.3</td><td>1437±24.0</td><td>1.431±0.015</td><td>1468±34.4</td><td>22.7± 0.31</td><td>1.984±0.040</td><td>13.0± 0.16</td></tr><tr><td>FAI T+N</td><td>2034±17.3</td><td>1436±17.6</td><td>1.416±0.017</td><td>1507±26.8</td><td>23.0± 0.61</td><td>1.987±0.061</td><td>12.9± 0.15</td></tr><tr><td>FAI T+CA</td><td>2058±15.6</td><td>1418±17.9</td><td>1.451±0.015</td><td>1467±51.9</td><td>23.3± 0.34</td><td>1.970±0.042</td><td>13.0± 0.15</td></tr><tr><td>FAI T+CA+N</td><td>2011±24.1</td><td>1413±18.8</td><td>1.429±0.012</td><td>1431±26.1</td><td>22.7± 0.36</td><td>2.018±0.045</td><td>12.9± 0.13</td></tr><tr><td>P-value</td><td>0.226</td><td>0.916</td><td>0.100</td><td>0.428</td><td>0.336</td><td>0.463</td><td>0.751</td></tr><tr><td>CV (%)</td><td>2.49</td><td>3.44</td><td>2.70</td><td>6.22</td><td>4.68</td><td>6.45</td><td>3.18</td></tr></tbody></table>

FI – Feed intake; WG – weight gain; FCR – feed conversion ratio; BW – body weight; BY – breast yield; LiY – liver yield; LeY – leg yield.

## Literature

BRUNSCHWILER, C., D. HEINE, S. KAPPELER, B. CONDE-PETIT, L. NISTRÖM, 2013: Direct measurement of rice bran lipase activity for inactivation kinetics and storage stability prediction. J. Cereal Sci. **58**, 272-277.

MUJAHID, A., I.U, HAQ, M. ASIF, A. GILANI, 2005: Effect of various processing techniques and different levels of antioxidant on stability of rice bran during storage. J. Sci. Food Agric. **85**, 847–85.

MUJAHID, A., I.U. HAQ, M. ASIF, A.H. GILANI, 2004: Effect of different levels of rice bran processed by various techniques on performance of broiler chicks. Br. Poult. Sci. **45**, 395-399.

ROSTAGNO, H.S., L.F.T. ALBINO, J.L. DONZELE, P.C. GOMES, R.F. OLIVEIRA, D.C. LOPES, A.S. FERREIRA, S.L.T. BARRETO, R.F. EUCLIDES, 2011: Tabelas Brasileiras para aves e suínos: composição de alimentos e exigências nutricionais. 252p.
P-152 (ID 141)

EFFECT OF ONION AND GRAPE SEED EXTRACTS ON MEAT QUALITY AND ANTIOXIDANT STATUS IN BROILER CHICKENS

K. Lipiński, Z. Antoszkiewicz, M. Mazur, J. Kaliniewicz and Z. Makowski

Dept. Animal Nutrition and Feed Science, University of Warmia and Mazury in Olsztyn, Oczapowskiego 5, 10-718 Olsztyn, Poland

Corresponding author: krzysztof.lipinski@uwm.edu.pl

**Summary**

A 34-day experiment was conducted on 80 day-old female broiler chickens (4 treatments, 10 replication, Ross 308) to determine the effect of vitamin E and natural polyphenols (onion and grape seed extracts, Proviox) on meat quality and antioxidant status. The birds received basal diets (starter, grower) without added vitamin E (a) or diets supplemented with vitamin E at 50/25 (b), 100/50 (c), 50/25 mg/kg and Proviox, 50/25 mg/kg (d). The following criteria were monitored: performance, carcass, meat quality and blood indicators (GPx, SOD, TAS and α-tocopherol). There were no differences in performance, dressing percentage, breast meat yield and chemical composition of meat. The experimental diets had no effect on the meat quality, with the exception of colour. The *b*\* value (yellowness) of meat significantly increased in the group with polyphenol supplementation (P≤0.05). As the supplemental vitamin E level increased, the serum α-tocopherol concentrations and SOD activity increased (P≤0.001). The chickens that received diets with the polyphenol supplement (d) were characterized by similar (SOD) or better (TAS, GPx) antioxidant status compared to the group fed diets only with the addition of vitamin E (b). The antioxidant status (TAS, GPx) was similar in groups c and d.

**Keywords:** polyphenols, vitamin E, meat quality, antioxidant status, broiler chickens

**Introduction**

Oxidative stress can cause a number of diseases, including cancer and degenerative diseases, since free radicals damage cells, and react with DNA, RNA, proteins and lipids (Lykkesfeldt and Svendsen, 2007; Voljč et al., 2011; Yin et al., 2013). The functions of the immune system become impaired. Endotoxins produced as a result of oxidative stress cause liver dysfunctions and defects in meat. Excessive oxidative stress is also a reason for problems related to reproduction and inferior productivity (Fellenberg and Speisky, 2006; Durand et al., 2013; Gobert et al., 2013). The above-mentioned phenomenon results from impaired mechanisms regulating the production of free radicals (diseases, environmental pollution and stress). Another oxidative stress inducing factor may involve nutritional factors such as unbalanced feed rations, poor quality feed fat, large amounts of polyunsaturated fatty acids, feed contaminated with mycotoxins, heavy metals, deficiency of natural antioxidants (vitamin E and C). The results of the study indicate that diets rich in natural antioxidants: vitamin A, C and E, carotenoids and polyphenols, can significantly affect the growth of reactive antioxidant status in the body, and thereby reduce the risk of certain diseases and favourably influence the performance. An interesting scientific problem is a possibility of the partial supplementation of vitamin E with preparations containing plant polyphenols, which can support (or "save") antioxidative functions of vitamin E (Surai 2014). Such opportunities have been confirmed in studies on pigs and poultry (Brenes et al., 2008; Frankič et al., 2011). The aim of the study was to determine the effect of natural polyphenols (Proviox) on the performance, carcass and meat quality, and on antioxidant and vitamin E status in broiler chickens.

**Material and Methods**

A 34-day experiment was conducted on 80 day-old female broiler chickens (4 treatments, 10 replication, Ross 308) to determine the effect of vitamin E and natural polyphenols (onion and grape seed extracts, Proviox) on meat quality and antioxidant status. The birds received basal diets (starter, grower) without added vitamin E (a) or diets supplemented with vitamin E at 50/25 (b), 100/50 (c), 50/25 mg/kg and Proviox, 50/25 mg/kg (d). Water and feed were provided ad libitum. Broilers were fed commercial starter and grower diets. The following criteria were monitored: performance (BW, FCR, European Broiler Index - EBI), carcass (carcass dressing percentage, breast muscle), meat quality (chemical composition, pH₁, pH₂₄, colour measurement (L, a, b), water holding capacity, drip loss) and blood indicators (glutathione peroxidase-GPx, superoxide dismutase-SOD activity, total antioxidant status-TAS and α-tocopherol concentrations). The analysis of chickens for fattening after slaughter was conducted on the 34th day of their lives. In total, 80 broilers (females) were slaughtered. Their gizzards, hearts and livers were dissected and the weight of each was de-
termed. Samples weighing 25 g were collected from the breast muscles. The composition was determined in the sample using the standard methods (AOAC 2005). Measurements of pH₁₅ and a 24h post-mortem of the breast meat were made (using Hanna Instruments 8314). Meat analyses were carried out using a colorimeter (Konica Minolta Chroma Meter), based on the CIELab colour system in which three parameters are determined: a* - redness; b* - yellowness, L* - lightness. Drip loss was determined using the suspension method according to Prangle et al. (1998). Water-holding capacity was determined according to Grau and Hamm (1952). To determine oxidative status, blood samples were collected from the broiler chickens in tubes with a lithium anticoagulant, and whole blood was used for the analysis. The content of TAS was determined using the Antioxidant assay kit (Cat. No CS0790), the activity of SOD was determined using SOD determination kit 19160, and GPx activity used assay Cellular Activity Kit Cat. No CGP1. The level of α-tocopherol was determined by reversed-phase high-performance liquid chromatography (HPLC) with a SHIMADZU chromatograph. The results were processed statistically by one-way ANOVA and Duncan's test. Arithmetic means (x), standard errors of the mean (SEM) and significance level (P) were determined. All calculations were performed using STATISTICA 10 software.

## Results

The inclusion of vitamin E and Proviox in the experimental diets increased body weight. On day 34 (the end of fattening), the group D birds (vitamin E+Proviox; 1,785 g) were characterized by the highest body weight. The noted difference reached approximately 120 g when compared to the control group of birds (1,662 g), but it was not statistically significant. The feed conversion ratio was not affected by the inclusion of vitamin E and Proviox. All birds remained in good health throughout the trial. There were no differences in dressing percentage, breast meat yield or the chemical composition of the meat. Acidity (pH), water-holding and drip loss were measured in muscle samples as parameters of meat quality. In comparison with the control group, no significant differences were evident in the chickens fed vitamin E-enriched diets. The experimental diets had no effect on meat quality, with the exception of colour. The b* value (yellowness) of meat significantly increased in the group with polyphenol supplementation (P≤0.05). Serum α-tocopherol concentrations were higher (P≤0.001) when vitamin E was added to the diets (Table 1). The addition of various levels of vitamin E and Proviox modified the TAS. For TAS, there were differences between the control (no vitamin E) and experimental groups. TAS serum increased more in chickens fed vitamin E with Proviox than in chickens fed only vitamin E. SOD serum (superoxide dismutase) values ranged from 22.59 to 72.47 enzyme units (U/ml). The highest SOD activities were observed in chickens fed a high level of vitamin E. GPx activity in the blood from chickens in the control group was significantly lower (P≤0.001) than in chickens receiving vitamin E. The highest GPx activities were observed in chickens fed vitamin E and Proviox and in those fed a higher level of vitamin E (P≤0.001).

<table><caption>**Table 1.** Effect of dietary vitamin E level and polyphenols supplementation on α-tocopherol (mg/ml serum) and TAS (mmol/L serum) concentrations, the activity of SOD (U/ml of serum) and GPx (U/ml of blood) in the serum and in blood broiler chickens at 34 days</caption><thead><tr><th rowspan="3">Specification</th><th colspan="5">Groups</th></tr><tr><th rowspan="2">K-</th><th rowspan="2">Vit. E 50/25</th><th colspan="2">Vit. E</th><th rowspan="2">SEM</th><th rowspan="2">P</th></tr><tr><th>Vit. E 100/50</th><th>50/25<br>Proviox<br>50/25</th></tr></thead><tbody><tr><td>α-tocopherol</td><td>0.32<sup>C</sup></td><td>1.10<sup>B</sup></td><td>1.33<sup>A</sup></td><td>1.05<sup>B</sup></td><td>0.064</td><td>&lt;0.001</td></tr><tr><td>TAS</td><td>0.26<sup>B</sup></td><td>0.28<sup>B</sup></td><td>0.34<sup>A</sup></td><td>0.33<sup>A</sup></td><td>0.007</td><td>&lt;0.001</td></tr><tr><td>SOD</td><td>22.59<sup>C</sup></td><td>43.73<sup>B</sup></td><td>72.47<sup>A</sup></td><td>48.10<sup>B</sup></td><td>2.980</td><td>&lt;0.001</td></tr><tr><td>GPx</td><td>30.51<sup>C</sup></td><td>40.25<sup>B</sup></td><td>68.87<sup>A</sup></td><td>69.20<sup>A</sup></td><td>2.863</td><td>&lt;0.001</td></tr></tbody></table>

A, B - P≤0.01
Conclusions

Based on the results, it can be concluded that the chickens that received diets with the polyphenol supplement (Proviox) were characterized by an antioxidant status similar (SOD) or superior (TAS, GPx) in comparison to the group fed with diets with moderate level of vitamin E. The antioxidant status (TAS, GPx) was similar for groups fed with diets with high level of vitamin E and with vitamin E and polyphenols supplementation. The experimental diets had no effect on the performance and meat quality, with the exception of colour.

Literature

BRENES A., A. VIVEROS, I. GONI, C. CENTENO, S.G. SÁYAGO-AYERDY, I. ARIJA, F. SAURA-CALIXTO, 2008: Effect of Grape pomace concentrate and vitamin E on digestibility of polyphenols and antioxidant activity in chickens. Poult. Sci., 87, 307-316.

DURAND D., M. DAMON, M. GOBERT, 2013: Oxidative stress in farm animals: general aspects. Cah. Nutr. Diet., 48, 218-224.53-70.

FELLENBERG M.A., H. SPEISKY, 2006: Antioxidants: their effects on broiler oxidative stress and its meat oxidative stability. World's Poult. Sci. J., 62-70.

FRANKIČ T., J. SALOBIR, 2011: *In vivo* antioxidant potential of Sweet chestnut (Castanea sativa Mill.) wood extract in young growing pigs exposed to *n*-3 PUFA-induced oxidative stress. J. Sci. Food Agric., 91, 1432-1439.

GOBERT M., M. DAMON, D. Durand, 2013: Oxidative stress and nutritional quality of animal products. Cah. Nutr. Diet., 48, 225-232.

LYKKESFELDT J., O. SVENDSEN, 2007: Oxidants and antioxidants in disease: Oxidative stress in farm animals. Vet. J., 2007, 173, 502-511.

SURAI P.F., 2014: Polyphenol compounds in the chicken/animal diet: from the past to the future. J. Anim. Physiol. Anim. Nutr., 98, 19-31;

VOLJČ M., T. FRANKIČ, A. LEVART, M. NEMEC, F. SALOBIR, 2011: Evaluation of different vitamin E recommendations and bioactivity of α-tocopherol isomers in broiler nutrition by measuring oxidative stress in vivo and the oxidative stability of meat. Poultry Sci., 90, 1478-1488.

YIN J., M.M. WU, H. XIAO, W.K. REN, J.L. DUAN, G. YANG, T.J. LI Y.L. YIN, 2014: Development of an antioxidant system after early weaning in piglets. J. Anim. Sci., 92:612-619.
P-153 (ID 190)

EFFECT OF TYPE OF CEREAL AND OAT HULLS ADDITION ON GASTROINTESTINAL MORPHOLOGY IN BROILERS ORALLY INFECTED WITH CAMPYLOBACTER JEJUNI

C. Millán<fnref n="1" />, A. Martín<fnref n="2" />, J. Mayot<fnref n="3" />, J. Sánchez<fnref n="1" />, and M. I. Gracia<fnref n="1" />

**Corresponding author:** mgracia@e-imasde.com

**Abstract**

An experiment was conducted within the EU-FP7 project CAMPYBRO for evaluating the effect of type of cereal and oat hulls (OH) addition on gastrointestinal (GIT) morphology in broilers. There were five treatments: corn [C], wheat [W] or barley [B], and the inclusion of 5% of OH in C and W diets. A total of 180 one-day-old Ross 308 broilers were divided into cages (3 birds/cage) and experimental treatments (12 cages/treatment). At 14 d, all broilers were orally gavaged with 100 µl of a solution containing 1x10⁵ cfu/ml of ST-45 C. jejuni. Campylobacter effect on GIT morphology could not be analysed since all birds were infected. On days 35 and 42, the performance parameters and the weight of GIT, empty BW (EBW), proventriculus (PRO), gizzard (GIZ), and the pH at PRO and GIZ were taken. Data were analysed by GLM procedure of SPSS (v.19.0). From 0-42d birds fed B ate more feed than birds fed C-OH. Age of animals significantly affected all the GIT parameters studied decreasing their proportion in relation to BW, GIT or EBW. Birds fed B showed the biggest GIT as %BW, and OH increased the GIT weight in C and W diets and the relative importance of the GIZ, but decreased it in the PRO. Dietary treatment did not affect the pH at GIZ, but OH increased the pH at the PRO, especially in W diet. It is concluded that type of diet modifies the GIT morphology.

**Keywords:** Campylobacter jejuni, cereals, oat hulls, broiler, gastrointestinal tract

**Introduction**

To date, several research have investigated beneficial effects of adding oat hulls (OH), or other types or fibrous fractions into the diet of broiler chickens (HETLAND et al., 2003). Moderate amounts of some fiber sources dilutes the diet but might improve the motility and function of the gastrointestinal tract (GIT) and, eventually benefit digestive physiology without compromising growth in broiler chickens (MATEOS et al., 2012). The objective of this study was to evaluate the effect of different cereal sourced and oat hulls (OH) addition on gastrointestinal morphology in broilers orally infected with Campylobacter jejuni.

**Material and Methods**

A total of 180 Ross 308 broilers (50% male and 50% female) from 1 to 42 days of age were used and allocated to the experimental treatments. There were five treatments applied from 1 to 42 d of age: corn (C), wheat (W) or barley (B), and the inclusion of 5% of OH in C and W diets. The trial was carried out in an Animal Biosafety Level 2 Unit for broilers. Broilers were housed in cages in groups of three. Mash (non-pelleted) feeds were fed ad libitum with no added coccidiostat, growth promoter or veterinary antibiotics. At 14 d of age, all broilers were orally gavaged with 100 µl of a solution containing 1 x 10⁵ CFU/ml of ST-45 C. jejuni strain. Performance was calculated by cage at 21 and 42 days. On days 35 and 42, twelve broilers per treatment were euthanized and the weight of the GIT, empty BW (EBW), proventriculus (PRO), gizzard (GIZ), and the pH at PRO and GIZ were taken. Data were analysed by GLM procedure (SPSS v.19.0). The model included the dietary treatment, the age and its interaction. A Tukey's test was used for mean comparison.

**Results**

There was no effect of dietary treatment on growth or feed conversion at any of the studied periods. From 0-21d, broilers fed wheat+OH showed higher feed intake that birds fed corn or wheat-based diets, showing the rest of treatments intermediate results. From 21-42d, broilers fed barley ate more feed than birds fed corn+OH and wheat+OH, showing the rest of treatments intermediate values. For the whole trial, broilers fed barley ate more feed than birds fed corn+OH (Table 1).

<footnote n="1">¹Imasde Agroalimentaria, S.L., Madrid, Spain</footnote>

<footnote n="2">²PROPOLLO, Madrid, Spain</footnote>

<footnote n="3">³FIA, Paris, France</footnote>
<table><caption>**Table 1.** Effect of dietary treatment on average daily gain (ADG, g), average daily feed intake (ADFI, g) and feed conversion ratio (FCR, g/g) from 0-21, 21-42 and 0-42 d of age</caption><thead><tr><th rowspan="2">Treatment</th><th colspan="3">0-21 d</th><th colspan="3">21-42 d</th><th colspan="3">0-42 d</th></tr><tr><th>ADG</th><th>ADFI</th><th>FCR</th><th>ADG</th><th>ADFI</th><th>FCR</th><th>ADG</th><th>ADFI</th><th>FCR</th></tr></thead><tbody><tr><td>Barley</td><td>31.7</td><td>64.1<sup>ab</sup></td><td>2.038</td><td>78.1</td><td>147.9<sup>a</sup></td><td>1.921</td><td>54.9</td><td>106.2<sup>a</sup></td><td>1.949</td></tr><tr><td>Corn</td><td>32.4</td><td>56.9<sup>b</sup></td><td>1.815</td><td>74.2</td><td>137.9<sup>ab</sup></td><td>1.897</td><td>53.8</td><td>98.3<sup>ab</sup></td><td>1.837</td></tr><tr><td>Corn + 5% OH</td><td>32.2</td><td>64.3<sup>ab</sup></td><td>2.013</td><td>69.8</td><td>120.6<sup>c</sup></td><td>1.745</td><td>51.1</td><td>92.7<sup>b</sup></td><td>1.817</td></tr><tr><td>Wheat</td><td>31.5</td><td>57.8<sup>b</sup></td><td>1.849</td><td>73.9</td><td>135.5<sup>ab</sup></td><td>1.854</td><td>53.3</td><td>97.7<sup>ab</sup></td><td>1.844</td></tr><tr><td>Wheat + 5% OH</td><td>28.6</td><td>69.5<sup>a</sup></td><td>2.075</td><td>76.0</td><td>130.6<sup>bc</sup></td><td>1.731</td><td>52.0</td><td>100.5<sup>ab</sup></td><td>1.941</td></tr><tr><td>SEM (n=12)</td><td>1.192</td><td>1.445</td><td>0.091</td><td>1.920</td><td>2.062</td><td>0.049</td><td>0.988</td><td>1.447</td><td>0.033</td></tr><tr><td>P</td><td>0.519</td><td><strong>0.001</strong></td><td>0.509</td><td>0.339</td><td><strong>0.001</strong></td><td>0.244</td><td>0.408</td><td><strong>0.001</strong></td><td>0.193</td></tr></tbody></table>

SEM = Standard error of mean. Different superscripts in same column are significant (a-c: P ≤ 0.05).

*Campylobacter* effect on GIT morphology could not be analysed since all birds were infected. Age of animals significantly affected to all the parameters studied showing, in general, bigger size of organs at 42d than at 35d, but lower importance expressed as percentage of BW, GIT, or EBW (Table 2).

Barley-based diets showed the biggest GIT in weight and in %BW, and consequently the lowest EBW, significant compared to corn and wheat diets (Table 3). Unusually, OH inclusion in corn and wheat diets decreased the relative importance of the proventriculus, compared to the cereals alone (4.14<sup>ab</sup>, 4.65<sup>a</sup>, 3.96<sup>b</sup>, 4.56<sup>a</sup> and 3.91<sup>b</sup> %BW for barley, corn, corn+OH, wheat and wheat+OH, respectively). As expected, OH inclusion in corn and wheat diets increased the relative importance of the gizzard compared to the cereals alone (2.87<sup>ab</sup>, 2.67<sup>bc</sup>, 3.14<sup>a</sup>, 2.47<sup>c</sup> and 2.89<sup>ab</sup> %BW for barley, corn, corn+OH, wheat and wheat+OH, respectively). This was a clear effect of long size particles of fiber, which stimulate the gizzard function and muscle development. Dietary treatment did not affect the pH at gizzard but, unexpectedly, OH increased the pH at the proventriculus, especially in wheat-based diets (3.69<sup>c</sup>, 3.72<sup>bc</sup>, 4.16<sup>ab</sup>, 3.70<sup>c</sup> and 4.26<sup>a</sup>, for barley, corn, corn+OH, wheat and wheat+OH, respectively).

## Conclusions

Under our experimental conditions, it is concluded that cereal type and quantity of fibre modifies the GIT morphology.

## Acknowledgements

This work has been done within the project CAMPYBRO, which has received funding from the European Union's Seventh Framework Programme for research, technological development and demonstration under grant agreement no 605835.

## Literature

HETLAND, H., B. SVIHUS, Å. KROGDAHL, 2003: Effects of oat hulls and wood shavings on digestion in broilers and layers fed diets based on whole or ground wheat. Br. Poult. Sci. **44**, 275-282.

MATEOS, G.G., E. JIMENEZ-MORENO, M.P. SERRANO, R. LAZARO, 2012: Poultry response to high elvels of dietary dietary fibre sources varying in physical and chemical characteristics. J. Appl. Poult. Res. **21**, 156-174.
<table><caption>**Table 2.** Effect of age, type of cereal and oat hulls (OH) inclusion (mean values at 35 and 42 d) on gastrointestinal tract (GIT) morphology and pH</caption><thead><tr><th rowspan="2"></th><th rowspan="2">BW, g</th><th colspan="2">GIT</th><th colspan="2">Empty BW</th><th colspan="4">Proventriculus</th></tr><tr><th>g</th><th>%BW</th><th>g</th><th>%BW</th><th>g</th><th>%GIT</th><th>%BW</th><th>%EBW</th></tr></thead><tbody><tr><td><strong>Age, d</strong></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>35d</td><td>1713</td><td>166.1</td><td>9.69</td><td>1547.3</td><td>90.31</td><td>6.95</td><td>4.23</td><td>0.41</td><td>0.45</td></tr><tr><td>42d</td><td>2261</td><td>208.3</td><td>9.20</td><td>2052.4</td><td>90.80</td><td>8.73</td><td>4.26</td><td>0.39</td><td>0.43</td></tr><tr><td>SEM (n=60)</td><td>27.3</td><td>3.82</td><td>0.135</td><td>24.87</td><td>0.135</td><td>0.185</td><td>0.094</td><td>0.007</td><td>0.008</td></tr><tr><td><strong>Treatment</strong></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>Barley</td><td>2060</td><td>210.7<sup>a</sup></td><td>10.26<sup>a</sup></td><td>1849.0</td><td>89.74<sup>b</sup></td><td>8.63<sup>a</sup></td><td>4.14<sup>ab</sup></td><td>0.42<sup>a</sup></td><td>0.47<sup>a</sup></td></tr><tr><td>Corn</td><td>1964</td><td>175.3<sup>b</sup></td><td>8.91<sup>b</sup></td><td>1788.7</td><td>91.09<sup>a</sup></td><td>8.17<sup>ab</sup></td><td>4.65<sup>a</sup></td><td>0.41<sup>ab</sup></td><td>0.45<sup>ab</sup></td></tr><tr><td>Corn + 5% OH</td><td>1938</td><td>182.6<sup>b</sup></td><td>9.45<sup>ab</sup></td><td>1755.5</td><td>90.55<sup>ab</sup></td><td>7.13<sup>b</sup></td><td>3.96<sup>b</sup></td><td>0.37<sup>b</sup></td><td>0.41<sup>b</sup></td></tr><tr><td>Wheat</td><td>2017</td><td>180.5<sup>b</sup></td><td>9.00<sup>b</sup></td><td>1836.8</td><td>91.00<sup>a</sup></td><td>8.04<sup>ab</sup></td><td>4.56<sup>a</sup></td><td>0.40<sup>ab</sup></td><td>0.44<sup>ab</sup></td></tr><tr><td>Wheat + 5% OH</td><td>1956</td><td>186.8<sup>b</sup></td><td>9.60<sup>ab</sup></td><td>1769.2</td><td>90.40<sup>ab</sup></td><td>7.25<sup>b</sup></td><td>3.91<sup>b</sup></td><td>0.37<sup>b</sup></td><td>0.41<sup>b</sup></td></tr><tr><td>SEM (n=24)</td><td>43.2</td><td>6.04</td><td>0.213</td><td>39.33</td><td>0.213</td><td>0.293</td><td>0.149</td><td>0.011</td><td>0.013</td></tr><tr><td><strong>Probability</strong></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>Age</td><td><strong>&lt;0.001</strong></td><td><strong>&lt;0.001</strong></td><td><strong>0.012</strong></td><td><strong>&lt;0.001</strong></td><td><strong>0.012</strong></td><td><strong>&lt;0.001</strong></td><td>0.867</td><td><strong>0.051</strong></td><td><strong>0.038</strong></td></tr><tr><td>Treatment</td><td>0.256</td><td><strong>0.001</strong></td><td><strong>&lt;0.001</strong></td><td>0.359</td><td><strong>&lt;0.001</strong></td><td><strong>0.001</strong></td><td><strong>0.001</strong></td><td><strong>0.002</strong></td><td><strong>0.002</strong></td></tr><tr><td>Age x Treatment</td><td>0.817</td><td>0.469</td><td>0.692</td><td>0.876</td><td>0.692</td><td>0.481</td><td>0.833</td><td>0.517</td><td>0.502</td></tr></tbody></table>

<table><thead><tr><th rowspan="2"></th><th colspan="4">Gizzard</th><th colspan="2">pH</th></tr><tr><th>g</th><th>%GIT</th><th>%BW</th><th>%EBW</th><th>Proventriculus</th><th>Gizzard</th></tr></thead><tbody><tr><td><strong>Age, d</strong></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>35d</td><td>50.45</td><td>30.44</td><td>2.95</td><td>3.27</td><td>3.90</td><td>3.36</td></tr><tr><td>42d</td><td>59.92</td><td>29.11</td><td>2.67</td><td>2.94</td><td>3.91</td><td>3.51</td></tr><tr><td>SEM (n=60)</td><td>1,203</td><td>0.458</td><td>0.054</td><td>0.062</td><td>0.072</td><td>0.063</td></tr><tr><td><strong>Treatment</strong></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>Barley</td><td>57.92<sup>ab</sup></td><td>27.85<sup>b</sup></td><td>2.87<sup>ab</sup></td><td>3.20<sup>ab</sup></td><td>3.69<sup>c</sup></td><td>3.50</td></tr><tr><td>Corn</td><td>52.29<sup>bc</sup></td><td>29.89<sup>b</sup></td><td>2.67<sup>bc</sup></td><td>2.93<sup>bc</sup></td><td>3.72<sup>bc</sup></td><td>3.23</td></tr><tr><td>Corn + 5% OH</td><td>60.54<sup>a</sup></td><td>33.57<sup>a</sup></td><td>3.14<sup>a</sup></td><td>3.47<sup>a</sup></td><td>4.16<sup>ab</sup></td><td>3.32</td></tr><tr><td>Wheat</td><td>49.08<sup>c</sup></td><td>27.39<sup>b</sup></td><td>2.47<sup>c</sup></td><td>2.72<sup>c</sup></td><td>3.70<sup>c</sup></td><td>3.56</td></tr><tr><td>Wheat + 5% OH</td><td>56.08<sup>abc</sup></td><td>30.19<sup>b</sup></td><td>2.89<sup>ab</sup></td><td>3.20<sup>ab</sup></td><td>4.26<sup>a</sup></td><td>3.55</td></tr><tr><td>SEM (n=24)</td><td>1.902</td><td>0.723</td><td>0.085</td><td>0.098</td><td>0.114</td><td>0.100</td></tr><tr><td><strong>Probability</strong></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>Age</td><td><strong>&lt;0.001</strong></td><td><strong>0.043</strong></td><td><strong>&lt;0.001</strong></td><td><strong>&lt;0.001</strong></td><td>0.952</td><td>0.087</td></tr><tr><td>Treatment</td><td><strong>&lt;0.001</strong></td><td><strong>&lt;0.001</strong></td><td><strong>&lt;0.001</strong></td><td><strong>&lt;0.001</strong></td><td><strong>&lt;0.001</strong></td><td><strong>0.070</strong></td></tr><tr><td>Age x Treatment</td><td>0.505</td><td>0.066</td><td>0.405</td><td>0.455</td><td>0.373</td><td>0.858</td></tr></tbody></table>

SEM = Standard error of mean. Different superscripts in same column are significant (a-c: P ≤ 0.05).
P-157 (ID 142)

EVALUATION OF CRIMPED KERNEL MAIZE SILAGE SUPPLEMENTATION ON BROILER PRODUCTION

Samir Ranjitkar<fnref n="1" />, Jette Søholm Petersen<fnref n="2" />, Ricarda M. Engberg<fnref n="1" />

**Corresponding author:** samir.ranjitkar@anis.au.dk

**Abstract**

A feeding trial was carried out to study the effect of dietary inclusion of crimped kernel maize silage (CKMS) on broiler production and nutrient digestibility. The CKMS had a low pH (4.2) and contained high amounts of lactic acid bacteria (10⁸ log cfu/g), and organic acids mainly lactic acid (212 mmol/kg) and acetic acid (96 mmol/kg). Four experimental diets were used: Wheat based feed (WBF), maize based feed (MBF), maize based feed supplemented with 15% CKMS (CKMS-15) and maize based feed supplemented with 30% CKMS (CKMS-30). The body weight at 37 days was highest in broilers fed MBF (2636g) and CKMS-15 (2683g) but lowest when CKMS-30 was fed (2497g). The low body weight in CKMS-30 might be explained by a lower apparent fat digestibility and increased feed particle size as indicated by a higher relative gizzard weight in this group. In general, the apparent digestibility of nutrients was lower when CKMS was fed. The feed dry matter intake was higher in all maize based groups compared to WBF. Further, the intake of water and the ratio of water:dry matter was lower in birds fed the three maize diets, and decreased further following addition of CKMS. Bird mortality decreased and litter quality improved when CKMS was fed, which was reflected by a lower foot pad lesion score in these two groups. With respect to production results and bird welfare, a dietary inclusion level of 15% CKMS showed promising results.

**Keywords:** poultry, nutrition, broiler, maize silage, fermented feed, production, welfare

**Introduction**

Maize is one of the most commonly used energy source in broiler feed throughout the world. In Denmark, maize is harvested late in November with an approximate water content of 40%. To provide storage stability and avoid costs related to artificial drying, kernel maize is crimped and ensiled (crimped kernel maize silage, CKMS). Similar to the fermentation of a compound feed (CANIBE AND JENSEN, 2012), CKMS is characterized by a low pH (pH 4) due to the presence of high concentration of lactic acid and acetic acid, and contains large numbers of lactic acid bacteria as a result of the fermentation process. All these factors, alone or in combination have been shown to reduce the colonization of enteropathogens, increase nutritive value of feed, and have positive influence on growth and performance. However, the fermentation of a compound feed has some disadvantages such as loss of synthetic amino acids and poor litter quality in poultry (ENGBERG et al., 2009). Therefore, the supplementation of a compound feed with separately fermented grains might be a better option. The aim of the present study was to investigate the influence of CKMS at inclusion levels of 15% and 30% on broiler performance and nutrient digestibility.

**Material and Methods**

A production trial was carried out with 736 Ross 308 male broiler chicks from 1-37 days. They were grouped into 4 treatments and were reared in pens (8 replicate pens) with free access of feed and water. The CKMS was prepared by ensiling the crimped kernel maize for approximately 8 weeks with addition of an organic acids (formic, propionic, and benzoic acids). Analysis of DM, pH and microbial counts were carried out on crimped maize before and after ensiling as described by ENGBERG et al. (2009). All groups received the same starter feed (1-7 days) containing 22.5% crude protein (CP) and 11.4 MJ metabolizable energy (ME) per kg. Four different grower diets were used in the period from 8 to 37 days. Group 1 received wheat based feed (WBF) with 59% wheat presenting the usual feeding regime in Denmark. Group 2 received a diet containing 58% maize (maize based feed, MBF). Group 3 and 4 received maize based feed supplemented with 15% CKMS (CKMS-15) and 30% CKMS (CKMS-30), respectively. The CKMS was replaced with maize on DM basis. Additionally, 15-30% and 10% whole wheat was added to WBF and all maize based diets, respectively. Whole wheat and CKMS were mixed with the pelleted compound feed before feeding. Gross energy, crude protein, fat, starch, sugar, ash, total non-starch polysaccharides (NSP), SCFA and lactic acid were determined (ENGBERG et al., 2009).

<footnote n="1">¹Department of Animal Science, Aarhus University, Tjele, Denmark</footnote>

<footnote n="2">²SEGES, Skejby, Aarhus, Denmark</footnote>
Body weight, feed-DM intake, feed conversion ratio (FCR), and water consumption of broilers were measured for each pen on day 1 and day 37. The litter DM content in each pen was analysed by freeze drying. Foot pads lesions of broilers (5 birds/pen) were scored on both feet at d 37. A digestibility study was carried out in metabolism cages from d 22-25. The DM, ash, fat, nitrogen and energy content in feed and excreta were analysed as stated above. Four birds from each pen were randomly killed by cervical dislocation on d 36 to measure the gizzard weight. Statistical analyses were performed using the GLM procedure of SAS® applying the following general model: Yi = μ + αi + εi, where Yi was the observed dependent variable, μ was the overall mean, αi was the effect of treatments and εi was the random error. Results are presented as least square means (LSMeans) and pooled standard error (SE). Probability values < 0.05 were considered as significant.

## Results

The pH and DM of CKMS was 4.2 and 53.5%, respectively. In CKMS, lactic acid bacteria (LAB) counts increased from 7.5 to 8.2 log cfu/g, while coliform bacteria, lactose negative bacteria, yeast and mould decreased below the detection limit (<3.0 log cfu/g) from 6-7 log cfu/g before ensiling. CKMS contained 212 mmol/kg lactic acid and 96 mmol/kg acetic acid. As a result, higher concentration of SCFA and lactic acid was determined in diets supplemented with CKMS (Table 1). The body weight of broilers in CKMS-15 was around 50g/bird and 100g/bird higher than birds receiving MBF and WBF, respectively (Table 2).

<table><caption>**Table 1** Composition of the starter and grower experimental diets</caption><thead><tr><th></th><th>WBF</th><th>MBF</th><th>CKMS-15</th><th>CKMS-30</th></tr></thead><tbody><tr><td>Dry matter (g/kg)</td><td>872.4</td><td>873.7</td><td>821.3</td><td>765.9</td></tr><tr><td><u>Per kg DM</u></td><td></td><td></td><td></td><td></td></tr><tr><td>Gross energy (MJ)</td><td>19.04</td><td>18.65</td><td>18.51</td><td>18.56</td></tr><tr><td>Crude protein (N x 6,25)</td><td>221.3</td><td>220.6</td><td>214.4</td><td>217.5</td></tr><tr><td>Starch</td><td>443.2</td><td>475.0</td><td>484.9</td><td>477.3</td></tr><tr><td>Total sugar</td><td>49.3</td><td>42.7</td><td>43.1</td><td>42.0</td></tr><tr><td>Total NSP</td><td>113</td><td>108</td><td>108</td><td>107</td></tr><tr><td>Fat</td><td>70.4</td><td>50.6</td><td>48.1</td><td>48.4</td></tr><tr><td>Ash</td><td>53.8</td><td>53.6</td><td>54.0</td><td>53.9</td></tr><tr><td><u>Per kg as fed</u></td><td></td><td></td><td></td><td></td></tr><tr><td>Short chain fatty acids</td><td></td><td></td><td></td><td></td></tr><tr><td>Acetic acid (mmol/kg)</td><td>6.2</td><td>6.4</td><td>13.8</td><td>18.5</td></tr><tr><td>Propionic acid (mmol/kg)</td><td>0.0</td><td>0.0</td><td>3.2</td><td>6.8</td></tr><tr><td>Benzoic acid (mmol/kg)</td><td>0.0</td><td>0.0</td><td>0.4</td><td>1.1</td></tr><tr><td>Lactic acid (mmol/kg)</td><td>2.1</td><td>2.0</td><td>26.7</td><td>66.3</td></tr></tbody></table>

Feed intake and FCR increased with the supplementation of CKMS. Both water consumption and total water-DM ratio was significantly lower in all maize based diets and further decreased with the supplementation of CKMS (Table 2). Apparent digestibility of fat (P=0.003) was lower in both groups fed CKMS whereas no difference was observed with respect to the digestibility of starch, ME and nitrogen retention. The relative gizzard weight of broilers (g/kg body weight) was higher in CKMS-30 than that of the other groups (P=0.001). Litter moisture (P=0.012) was lower in CKMS-30 (34.6%) and CKMS-15 (38.7%) compared to WBF (45.4%) and MBF (41.2%). Likewise, foot pad score was significantly lower (P<0.001) in CKMS-15 (3.4) and CKMS-30 (3.1) as compared to WBF (44.4) and MBF (28.8). Mortality of broilers was significantly higher (P<0.001) in WBF (7.6%) and MBF (4.9%) than in CKMS-15 (1.1%), whereas no mortality was observed in the CKMS-30.
<table><caption>**Table 2** Body weight, feed-DM intake, FCR, water consumption and total water-DM ratio of broilers on day 37</caption><thead><tr><th rowspan="2">Item</th><th rowspan="2">Day</th><th colspan="4">Diets</th><th rowspan="2">SE</th><th rowspan="2">P value</th></tr><tr><th>WBF<sup>1</sup></th><th>MBF<sup>2</sup></th><th>CKMS-15<sup>3</sup></th><th>CKMS30<sup>4</sup></th></tr></thead><tbody><tr><td>Body weight (g)</td><td>37</td><td>2574<sup>bc</sup></td><td>2636<sup>ab</sup></td><td>2683<sup>a</sup></td><td>2497<sup>c</sup></td><td>36.82</td><td>0.008</td></tr><tr><td>Feed DM consumption (g)</td><td>37</td><td>3858<sup>b</sup></td><td>4044<sup>ab</sup></td><td>4349<sup>a</sup></td><td>4296<sup>a</sup></td><td>103.13</td><td>0.007</td></tr><tr><td>Feed conversion ratio</td><td>37</td><td>1.50<sup>b</sup></td><td>1.53<sup>b</sup></td><td>1.62<sup>ab</sup></td><td>1.72<sup>a</sup></td><td>0.05</td><td>0.005</td></tr><tr><td>Water consumption (ml)</td><td>37</td><td>6118<sup>a</sup></td><td>5632<sup>b</sup></td><td>5693<sup>b</sup></td><td>5291<sup>c</sup></td><td>93.59</td><td>&lt;0.001</td></tr><tr><td>Total water-DM ratio (ml water/g feed DM)*</td><td>37</td><td>1.68<sup>a</sup></td><td>1.49<sup>b</sup></td><td>1.47<sup>b</sup></td><td>1.44<sup>b</sup></td><td>0.04</td><td>0.001</td></tr></tbody></table>

## Conclusions

The dietary inclusion of 15% CKMS in broilers feed resulted in increased body weight. However, broilers in CKMS-30 showed the lowest body weight which was associated with an increased feed particle size indicated by the higher giz-zard weight and reduced fat digestibility in this group (data not shown). Lower intake of water and total water-DM ratio (Table 2) had contributed to an improved litter quality which was reflected by the lower frequency of foot pad lesions in broilers fed CKMS-15 and CKMS-30, indicating a positive attribute from animal welfare perspective. Soluble NSP has been associated with increased water intake (FRANCESCH AND BRUFAU, 2004), and therefore presence of high NSP in WBF compared to maize (Table 1) could be a possible explanation. Furthermore, combination of improved litter condition along with higher acidification of the upper digestive tract related to higher concentration of organic acids in feed (HERES et al., 2004) conataining CKMS-15 and CKMS-30 (Table 1) might have resulted lower mortality in broilers fed CKMS.

## Literature

CANIBE, N., B. B. JENSEN, 2012: Fermented liquid feed-Microbial and nutritional aspects and impact on enteric diseases in pigs. Anim. Feed Sci. Technol. **173**, 17-40.

ENGBERG, R. M., M. HAMMERSHØJ, N. JOHANSEN, M. ABOUSEKKEN, S. STEENFELDT, B. B. JENSEN, 2009: Fermented feed for laying hens: effects on egg production, egg quality, plumage condition and composition and activity of the intestinal microflora. Br. Poult. Sci. **50**, 228-239.

FRANCESCH, M., J. BRUFAU, 2004: Nutritional factors affecting excreta/litter moisture and quality. World's Poult Sci J. **60**, 64-75.

HERES, L., B. ENGEL, H. A. P. URLINGS, J. A. WAGENAAR, F. VAN KNAPEN, 2004: Effect of acidified feed on susceptibility of broiler chickens to intestinal infection by Campylobacter and Salmonella. Vet Microbiol. **99**, 259-267.

<footnote><sup>a-c</sup>LSMEANS within the same row with different superscripts differ significantly (P < 0.05); *Including drinking water and dietary water</footnote>
P-158 (ID 126)

# EFFECT OF HOP CONE SUPPLEMENTATION ON OXIDATIVE STRESS AND OXIDATIVE STABILITY OF FRESH AND STORED MEAT IN BROILERS

V. Rezar, A. Levart, and J. Salobir

Dept. of Anim. Science, Biotechnical Faculty, University of Ljubljana, Slovenia

Corresponding author: janez.salobir@bf.uni-lj.si

## Abstract

Broilers are frequently exposed to oxidative stress which can lead to reduced health status, performance and stability of meat. Consumer concerns about feed additives led to attempts to find natural bio-efficient antioxidants for animal nutrition. Hops (*Humulus lupulus* L.), are rich in phenols with antioxidant activity (xanthohumol, isoxanthohumol). The study examined the effects of hop cone supplementation on oxidative stress *in vivo* and oxidative stability of meat in broilers. 84 one-day-old male broiler chickens were housed in six floor pens and divided into 3 groups of 28. All groups received a high-PUFA diet unsupplemented or supplemented as follows: no supplementation (C), 0.9 g hops/kg (H1) and 3.6 g hops/kg (H2). Oxidative stress *in vivo* was studied by measuring the rate of lymphocyte DNA damage, malondialdehyde (MDA) concentration in plasma and breast muscle and by total plasma antioxidant capacity of lipid- and water-soluble compounds. Oxidative stability of meat was studied by measuring MDA in fresh and stored (refrigerated, frozen) breast meat. Both hop concentrations reduced the rate of blood lymphocyte DNA damage. Higher hop supplementation induced MDA formation and reduced plasma antioxidant activity of lipid-soluble compounds. In H2 group an increased MDA formation was observed also. An effect on other parameters of meat quality was not observed. Further studies are suggested to confirm the results and clarify the possible use of hops in broiler nutrition.

**Keywords:** hops, antioxidants, PUFA, oxidative stress, broilers

## Introduction

In broilers, oxidative stress may occur as a consequence of nutrition such as high amount of dietary polyunsaturated fatty acids (PUFA), high environmental temperatures and also several pathological conditions. Lipid oxidation also has negative effects on meat quality and often determines the shelf life of meat and meat products. The high intake of PUFA is a very important factor that increases the requirement for antioxidants. Recent attempts to find novel bio-efficient antioxidants have focused particularly on natural antioxidants, in order to satisfy consumer concerns over the safety and toxicity of food additives. Hops (*Humulus lupulus* L.) are known as an excellent source of polyphenolic antioxidants including proanthocyanidins, flavanols, phenolic acids, stilbenes, flavonols and prenylated flavonoids. In addition, hop oil and hop acids have been reported as potent antioxidants (Yamaguchi et al., 2009). Much of attention has gone to the polyphenolic content of hops, and specific compounds, such as xanthohumol and 8-prenylnaringenin, have been identified as multipotent bioactive compounds. The objective of this study was to demonstrate the potential effectiveness of hops in terms of animal performance, antioxidant capacity *in vivo* and of oxidative stability of broiler meat under various storage conditions.

## Material and Methods

84 one-day-old male Ross 308 broiler chickens were caged in wire-floored cages. The diets were formulated according to Ross 308 broiler nutrition specifications. All groups received a high-PUFA diet containing 7.5 % linseed oil, supplemented with: no supplementation (C), supplemented with 0.9 g of hops/kg (H1) and with 3.6 g of hops/kg (H2). Hops were provided from Slovenian institute for hop research and brewing. After 37 days of treatment, the birds were killed. Blood and breast muscle samples from each bird were collected for analysis. Lymphocytes were isolated from the fresh blood samples. The comet assay was performed according to the standard method with slight modifications as described by Rezar et al. (2003). The concentrations of MDA in blood plasma and meat samples were determined by HPLC. Meat was exposed to different treatments: stored at -70°C till analysis (raw meat), refrigerated for 6 days at 4°C and then frozen for 3 months at -20°C, and refrigerated at -20°C for 3 months. Plasma antioxidative capacity of lipid-soluble substances (ACL) and water-soluble substances (ACW) were determined by photochemiluminescence detection using the Photochem assay (Analytik Jena, Leipzig, Germany). The animal ethics committee of the Veterinary Administration of the Republic of Slovenia approved the study protocol. Data were analysed using the GLM procedure of the SAS/STAT module (SAS 8e, 2000; SAS Institute Inc., Cary, NC). Significance was considered established at *P* < 0.05.
## Results

Oxidative stress can occur from providing large amounts of high-PUFA vegetable oils to reach the high energy requirements of poultry or when creating poultry products with improved nutritional quality (products rich in n-3 PUFA). The results from literature show, that bitter acids of hop cones and their analogues have antioxidant activity, potent DPPH radical scavenging activity and lipid peroxidation inhibitory activity (Tahashira et al., 1995). Plasma malondialdehyde (MDA) was measured as biomarker for oxidative stress *in vivo* (Nielsen et al., 1997). In comparison with C group, the supplement of 3.6 g/kg of hops in the feed significantly increased MDA concentration in plasma, which indicates that the highest level of hops supplementation increased oxidative stress. Oxidative damage to DNA has also been identified as a useful index of oxidative stress. In Table 1, DNA damage in blood lymphocytes is presented as percentage of DNA in the head of the comet. Results of the comet assay indicate that supplementation of 3.6 g hops/kg of feed has protective effect on lymphocyte's DNA due to decreased DNA fragmentation.

<table><caption>**Table 1.** Concentration of MDA in plasma (nmol/ml), lymphocyte DNA damage (expressed as % DNA in the head of the comet) and antioxidative capacity of water (ACW) and lipid (ACL) soluble antioxidants of chicken's plasma in different experimental groups</caption><thead><tr><th>Parameter/Group</th><th>C</th><th>H1</th><th>H2</th><th>P-value</th></tr></thead><tbody><tr><td>MDA in plasma (nmol/ml)</td><td>1.00 <sup>a</sup></td><td>2.06 <sup>a</sup></td><td>13.29 <sup>b</sup></td><td>&lt; 0.001</td></tr><tr><td>% DNA in head</td><td>74.96 <sup>a</sup></td><td>78.12 <sup>ab</sup></td><td>84.12 <sup>b</sup></td><td>0.0063</td></tr><tr><td>ACW (nmol AA* equiv/ml)</td><td>73.81</td><td>71.48</td><td>69.95</td><td>0.877</td></tr><tr><td>ACL (nmol trolox equiv/ml)</td><td>129.42 <sup>a</sup></td><td>125.96 <sup>a</sup></td><td>110.90 <sup>b</sup></td><td>0.0031</td></tr></tbody></table>

LS-means (least square means) in the same row without the same superscript (abc) differ significantly, *P* < 0.05

*AA – ascorbic acid

A hop is known to have antioxidant properties, therefore we also measured antioxidant capacity of lipid (ACL) and water (ACW) soluble antioxidants in chicken plasma samples to assess oxidative stress. As shown in Table 1, concentrations of ACW were similar between the groups, but ACL content were significantly lower in the group supplemented with 3.6 g hops/kg in comparison to the group C, which indicates that lipid soluble antioxidants were consumed as lipid oxidation progressed, which is in concordance with higher concentrations of plasma MDA.

Some previous studies showed that a diet rich in PUFA increased MDA levels in fresh and also in meat, stored under different conditions (Botsoglou et al., 2003). MDA concentration in raw meat was statistically higher in the group H2 in comparison the group C, as was in blood plasma. The MDA levels increased when breast meat was refrigerated for 6 days and then frozen for 3 months at -20 °C, but we did not find statistical differences between the groups.

<table><caption>**Table 2.** Concentration of MDA (nmol/g) in fresh and in different storage samples of breast meat</caption><thead><tr><th>Treatment/Group</th><th>C</th><th>H1</th><th>H2</th><th>P-value</th></tr></thead><tbody><tr><td>Raw meat</td><td>17.64 <sup>a</sup></td><td>21.88 <sup>ab</sup></td><td>26.36 <sup>b</sup></td><td>0.0523</td></tr><tr><td>Frozen for 3 months at -20°C</td><td>15.93</td><td>13.93</td><td>19.37</td><td>0.5908</td></tr><tr><td>Refrigerated for 6 days at 4°C than frozen for 3 months at -20°C</td><td>25.46</td><td>27.71</td><td>34.42</td><td>0.3493</td></tr></tbody></table>

LS-means (least square means) in the same row without the same superscript (abc) differ significantly, *P* < 0.05

## Conclusions

The results of the present study demonstrate the inconsistent effects of hop supplementation in two different concentrations on oxidative status of broilers and on the quality of fresh and stored broiler meat. The results of the experiment show, that supplementation of hops in concentration of 3.6 g/kg of feed have a negative effect and increased lipid peroxidation in chickens, measured as MDA concentration in the plasma. At the same time, hops had a protective effect on DNA of lymphocytes, increasing percentage of un-fragmented DNA. The rate of lipid oxidation in fresh meat, measured as formation of secondary product of lipid oxidation (MDA) demonstrates the detrimental effect of hops supplementation. Further studies are needed to confirm the results of the present study.
## Literature

BOTSOGLOU, N.A., S.H. GRIGOROPOULOU, E. BOTSOGLOU, A. GOVARIS, G. PAPAGEORGIOU, 2003. The effect of dietary oregano essential oil and α-tocopheryl acetate on lipid oxidation in raw and cooked turkey during refrigerated storage. Meat Sci. **65**: 1193–1200.

NIELSEN, F., B.B. MIKKELSEN, J.B. NIELSEN, H.R. ANDERSEN, P. GRANDJEAN, 1997. Plasma malondialdehyde as biomarker for oxidative stress: reference interval and effects of life-style factors. Clin. Chem. **43**: 1209–1214

REZAR, V., T. PAJK, R. MARINŠEK LOGAR, V. JEŠE JANEŽIČ, K. SALOBIR, A. OREŠNIK, 2003. Wheat bran and oat bran effectively reduce oxidative stress induced by high-fat diets in pigs. Ann. Nutr. Metab. **47**: 78–84.

TAGASHIRA, M., M. WATANABE, N. UEMITSU, 1995. Antioxidative activity of hop bitter acids and their analogues. Biosci. Biotechnol. Biochem. **59**, 4, 740–742.

YAMAGUCHI, N., K. SATOH-YAMAGUCHI, O. MITSUNORI, 2009: *In vitro* evaluation of antibacterial, anticollagenase, and antioxidant activities of hop components (*Humulus lupulus*) addressing acne vulgaris. Phytomed., **16**, 4, 369–376.
P-159 (ID 4)

EFFECT OF REPLACING SOY IN BROILER DIETS ON PERFORMANCE, CARCASS YIELD AND COST OF PRODUCTION

M. Ritter<fnref n="1" />, D. Albiker<fnref n="2" /> and P. Spring<fnref n="1" />

**Corresponding author:** peter.spring@bfh.ch

**Abstract**

The study aimed to evaluate the effects of replacing soy bean meal (SBM) in broiler diets on production parameters. A randomized trial (36 d) was conducted using 20 pens (20 m<fnref n="2" />, 280 Ross PM3 birds). A starter and grower diet (12.6/13.0 MJ ME, 210/200 g CP, 12.8/12.2 g lysine) were fed as crumbs. The diets contained either a high level of SBM (HS; starter/grower: 28.7/26.8% SBM and 2.5/2.5% rape seed meal (RSM)), a medium level of SBM (MS; starter/finisher: 15.0/15.0% SBM, 15.0/15.0% peas, 5.0/5.0% RSM, 5.0/5.0% sun flower meal (SFM), 2.8/2.0% potato protein (PP) or no SBM (NS; starter/grower: 25.0/25.0% peas, 5.0/5.0% RSM, 10.0/10.0% SFM, 5.3/4.5% PP, 4.0/4.0% corn gluten). Treatments were HS/HS, MS/MS, MS/NS, NS/NS for starter/finisher diets. Performance parameters, litter quality, hock burns (HB) and foot pad lesions (FPL) were recorded and analysed by using ANOVA and Tukey-Kramer.

Feed intake for HS/HS, MS/MS, MS/NS, NS/NS was 3438ª g, 3481ab g, 3574b g, 3580b g (P, 0.003). Body weight (Ø 2234 g), FCR (Ø 1.57) and mortality (Ø 2.14%) did not differ between treatments. The ratio of water/feed intake was highest in HS/HS. This led to lower litter quality and significantly higher scores for FPL compared to the other treatments. Carcass yield (Ø 69.50%) and composition did not differ between treatments. Feed ingredient costs were 5% higher for NS/NS compared to HS/HS. Overall performance was comparable between treatments, however, SBM led to significantly higher occurrence of FPL.

**Keywords:** Soy, protein alternatives, broiler, performance

**Introduction**

Markets and politics demand a more sustainable food production system. In the sustainability discussion soy production and trade is often a controversial issue. Even though 80 - 90 % of soy imported into Switzerland fulfills high standards concerning the sustainability of production, the acceptance for soy imports is missing by part of the consumers. Furthermore, the price for GMO-free soy has risen over the last years.

The objective of the present study was to evaluate the effect of replacing soy bean meal (SBM) in broiler diets on performance, carcass quality and production cost.

**Material and Methods**

The trial involved four dietary treatments and 5600 as hatched Ross PM3 birds. The birds (280 per pen) were randomly assigned to 20 pens of 20 m<fnref n="2" /> each. The trial lasted 36 days. Barn climate was based on the commercial standards for the breed. A starter diet (12.6 MJ ME, 210 g CP, 12.8 g Lysin, 9.0 g Ca und 6.5 g P) and a grower diet (13.0 MJ ME, 200 g CP, 12.2 g w, 7.5 g Ca und 5.5 g P) were fed as crumbs *ad libitum*. The diets contained either a high level of SBM (HS), a medium level of SBM (MS) or no SBM (NS) (Table 1). Treatments were HS/HS, MS/MS, MS/NS, NS/NS for starter/grower diets.

<table><caption>**Table 1.** Percentage of protein rich ingredients in the different diets (% in starter and grower)</caption><thead><tr><th>Protein rich ingredients</th><th>HS</th><th>MS</th><th>OS</th></tr></thead><tbody><tr><td>Soy, %</td><td>28.7/26.8</td><td>15.0/15.0</td><td>0/0</td></tr><tr><td>Peas, %</td><td>0/0</td><td>15.0/15.0</td><td>25.0/25.0</td></tr><tr><td>Rape cake, %</td><td>2.5/2.5</td><td>5.0/5.0</td><td>5.0/5.0</td></tr><tr><td>Sunflower meal, %</td><td>0/0</td><td>5.0/5.0</td><td>10.0/10.0</td></tr><tr><td>Potato protein, %</td><td>0/0</td><td>2.8/2.0</td><td>5.3/4.5</td></tr><tr><td>Corn gluten, %</td><td>0/0</td><td>0/0</td><td>4.0/4.0</td></tr></tbody></table>

<footnote n="1">¹ Berne University of Applied Sciences, HAFL-Agriculture, 3052 Zollikofen, Switzerland</footnote>

<footnote n="2">² Aviforum Foundation, 3052 Zollikofen, Switzerland</footnote>
Feed structure and composition were analyzed. Standard performance parameters, water intake, litter quality (% crusted surface), foot pad lesions and hock burns were recorded (0 = no change; 3 = strong change). In addition, carcass grading and composition were evaluated. Data were analyzed by ANOVA and means compared with the test of Tukey-Kramer.

## Results

Feed composition was in agreement with calculated values and feed structure was comparable between treatments (data not shown).

Treatments with no SBM in the grower diet led to a significant higher feed intake compared to HS/HS. Body weight, FCR and mortality were not affected by treatments. EBI was between 363.4 and 377.2 (Table 2).

<table><caption>**Table 2.** Effect of protein rich ingredients on performance of ROSS PM3 birds</caption><thead><tr><th>Parameters</th><th>HS/HS</th><th>MS/MS</th><th>MS/OS</th><th>OS/OS</th><th>SEM</th><th>P</th></tr></thead><tbody><tr><td>Feed intake</td><td>3438<sup>a</sup></td><td>3481<sup>ab</sup></td><td>3574<sup>b</sup></td><td>3580<sup>b</sup></td><td>26.667</td><td>0.003</td></tr><tr><td>Body weight, g</td><td>2219</td><td>2214</td><td>2229</td><td>2274</td><td>33.243</td><td>0.571</td></tr><tr><td>FCR g/g</td><td>1.549</td><td>1.572</td><td>1.606</td><td>1.574</td><td>0.015</td><td>0.086</td></tr><tr><td>Mortality, %</td><td>2.07</td><td>2.07</td><td>2.07</td><td>2.36</td><td>0.202</td><td>0.890</td></tr><tr><td>EBI</td><td>374.4</td><td>368.4</td><td>363.4</td><td>377.2</td><td>8.672</td><td>0.683</td></tr></tbody></table>

The ratio of water/feed intake was highest in HS/HS (Table 3). This led to lower litter quality (92% of surface crusted in HS/HS pens) and significantly higher scores for FPL compared to the other treatments. HS/HS also showed the highest average score for HB (Table 3).

<table><caption>**Table 3.** Effect of protein rich ingredients on water to feed intake, foot pad lesions and hock burns of ROSS PM3 birds</caption><thead><tr><th>Parameter</th><th>HS/HS</th><th>MS/MS</th><th>MS/OS</th><th>NS/OS</th><th>SEM</th><th>P</th></tr></thead><tbody><tr><td>g water / g feed</td><td>1.935<sup>a</sup></td><td>1.847<sup>ab</sup></td><td>1.693<sup>b</sup></td><td>1.790<sup>ab</sup></td><td>0.049</td><td>0.023</td></tr><tr><td>Foot pad lesion*</td><td>0.418<sup>a</sup></td><td>0.132<sup>b</sup></td><td>0.128<sup>b</sup></td><td>0.094<sup>b</sup></td><td>0.052</td><td>0.002</td></tr><tr><td>Hock burn*</td><td>0.340<sup>a</sup></td><td>0.110<sup>b</sup></td><td>0.080<sup>b</sup></td><td>0.140<sup>ab</sup></td><td>0.052</td><td>0.011</td></tr></tbody></table>

All treatments achieved excellent carcass quality with over 97.5% grade 1. Average carcass yield was 69.50% and was not affected by treatment. Dietary protein source did not influence carcass composition (Table 4).

<table><caption>**Table 4.** Effect of protein rich ingredients on carcass composition of ROSS PM3 birds</caption><thead><tr><th>Parameter</th><th>HS/HS</th><th>MS/MS</th><th>MS/OS</th><th>NS/OS</th><th>SEM</th><th>P</th></tr></thead><tbody><tr><td>Breast, %</td><td>28.05</td><td>27.45</td><td>27.95</td><td>27.35</td><td>0.0032</td><td>0.352</td></tr><tr><td>Leg, %</td><td>30.50</td><td>30.15</td><td>29.95</td><td>30.60</td><td>0.0026</td><td>0.373</td></tr><tr><td>Wing, %</td><td>10.30</td><td>10.15</td><td>10.25</td><td>10.30</td><td>0.0010</td><td>0.878</td></tr><tr><td>Rest, %</td><td>31.15</td><td>32.00</td><td>31.90</td><td>31.65</td><td>0.0029</td><td>0.100</td></tr></tbody></table>

## Conclusions

Diets with no SBM led to a significantly higher feed intake compared to diets with SBM. However, weight gain and FCR were not affected by diet composition. Overall performance of SBM-free diets was comparable to the standard commercial diet with SBM. Dietary composition did also not influence carcass composition. Carcass composition has been shown to change with suboptimal protein nutrition (SUMMERS ET AL. 1992; FARIA FILHO ET AL. 2005). The lack of difference indicates that all birds received adequate and comparable protein and amino acid supply.

SBM-free diets led to significantly lower water to feed intake. This could partially be explained by the lower potassium concentration in diets without SBM (VIEIRA & LIMA 2005; SMITH ET AL. 2000). The lower water intake led to healthier skin on the legs. Both foot pad lesions and hock burns were lower in diets with no SBM. This is in agreement with TUCKER and WALKER (1999) who also reported a link between litter quality and the occurrence of FPL and HB.

SBM could be replaced with little change in performance and a significant reduction in FPL and hock burns. However, feed ingredient cost were 5% higher for diets without SBM compared to diets with high inclusion of SBM. In the market,

<footnote>*day 35 Score 0 – 3 (0 = no change, 3 = strong change)</footnote>
not all alternative protein sources used in the trial (particularly potato protein) are available in sufficient amounts to switch a larger market segment to diets without SBM. Therefore, SBM-free diets could only play a role in niche markets where the extra cost can be passed on to the customer.

## Literature

FARIA FILHO, D. E., P. S. ROSA, B. S. VIEIRA, M. MACARI, R. L. FURLAN, 2005: Protein levels and environmental temperature effects on carcass characteristics, performance, and nitrogen excretion of broiler chickens from 7 to 21 days of age. Revista Brasileira de Ciência Avícola, 7. Jg., Nr. 4, p. 247-253.

SMITH, A., S. P. ROSE, R. G. WELLS, V. PIRGOZLIEV, 2000: Effect of excess dietary sodium, potassium, calcium and phosphorus on excreta moisture of laying hens. British Poultry Science, 2000, 41. Jg., Nr. 5, p. 598-607.

SUMMERS, J. D., D. SPRATT, J. L. ATKINSON, 1992: Broiler weight gain and carcass composition when fed diets varying in amino acid balance, dietary energy, and protein level. Poultry Science, 1992, 71. Jg., Nr. 2, p. 263-273.

TUCKER, S.A. and WALKER, A.W. 1999: Hock Burn in Broilers. In *Recent Developments in Poultry Nutrition* 2. Wise-man, J and Garnsworthy, P C. (Eds). Nottingham University Press, Nottingham. p. 107-122.

VIEIRA, S. L., I. L. LIMA, 2005: Live performance, water intake and excreta characteristics of broilers fed all vegetable diets based on corn and soybean meal. International Journal of Poultry Science, 2005, 4. Jg., Nr. 6, p. 365-368.
P-161 (ID 257)

GLYCEROL-A GREAT POTENTIAL SOURCE OF ENERGY FOR LAYER

Y. Ruangpanitand and P. Sriboonyong

Department of Animal Science, Kasetsart University, KamphaengSaen Campus, NakhonPathom, 73140 Thailand

Corresponding author: minicooper301@gmail.com

Abstract

The aim of the study was to investigate the use of purified glycerol from biodiesel production as energy feed-stuff for layer. Purified glycerol is used for drug and cosmetic production. It is also suitable for food and animal feed. Purified glycerol contains 99%wt up for glycerol content, less than 0.01%wt of methanol, 3842-4118 kcal/kg of gross energy and 3,352 kcal/kg ME for poultry. Therefore, the purified glycerol has a high potential as energy feed stuff for domestic animals. The study on productive performance was conducted to determine the use of purified glycerol in layer diet. A total of 480, 24 week of age, Lohmann Brown were divided into 4 treatments including, corn-soy basal diet (CS), CS with 2.5% of glycerol, CS with 5.0% of glycerol and CS with 7.5% of glycerol, respectively. Each treatment consisted of 10 replications with 12 birds each. All diets were isocaloric and isonitrogenous (2,750 kcal/kg, 17.5%CP). Birds were raised in an evaporative system for 4 periods, 28 days per period. There was no significant difference in feed intake, hen house production, hen day production and egg weight in each period. The result revealed that purified glycerol can be used in layer diet at 7.5% without a negative effect on productive performance.

**Keywords:** Glycerol, gross energy, metabolizable energy, energy source

Introduction

Nowadays, the price of oil is dramatically increasing. Thailand has promoted the alternative energy, biodiesel, to replace using fossil fuel. The raw materials for biodiesel production are mainly palm oil, and palm wax. The other materials used are cooking oil, animal fat and plant oil. Its chemical structure is that of fatty acid alkyl ester (FAAE). Biodiesel as a fuel gives much lower toxic air emissions than fossil fuel. In addition, it gives cleaner burning and has less sulfur content, and thus reducing emission. Because of its origin from renewable resources, it is more likely that it competes with petroleum products in future (HAYYAN et al., 2010). Commercially, biodiesel is produced by transesterification of triglycerides which are the main ingredients of biological origin oils in the presence of an alcohol (e.g. methanol, ethanol) and a catalyst (e.g. alkali, acid) with glycerol or glycerine as a major by-product (MA and HANNA, 1999). In recent years, increasing of biodiesel production has been associated with increasing glycerol production. Biodiesel production process produces approximately 10% of crude glycerol (Anonymous, 2007). The crude glycerol can be purified into purified glycerol for 30-45%. This depends on the quality of the raw materials. Purified glycerol is commonly used for drug and cosmetic production. It is also suitable for food and animal feed. The aim of the study was to investigate the use of purified glycerol from biodiesel production as energy feed-stuff for layer.

Materials and Methods

A total of 480, 24 week of age, Lohmann Brown were divided into 4 treatments including, corn-soy basal diet (CS), CS with 2.5, 5.0% and 7.5% of glycerol, respectively. Each treatment consisted of 10 replications with 12 birds each. Each replication contain 4 cages (20 cm × 40 cm × 36 cm per cage) with 3 hens per cage at light at 16 h light per day. Feed in mash form and water were provided *ad libitum*. All diets were isocaloric and isonitrogenous (2,750 kcal/kg ME, 17.5%CP). Birds were raised in an evaporative system for 4 periods with 28 days per period. The ingredients and chemical composition of the diet are presented in Table 1. Laying hens were weighed individually at the beginning and at the end of the experiment. Mortality was recorded as it occurred. Eggs were collected daily and egg production was expressed on a hen-day and hen-house basis. All the eggs laid were collected and weighed to determine the egg weight. Feed intake was weekly recorded then calculated as g per hen per day and expressed in each period.
<table><caption>**Table 1.** Ingredients and chemical composition of the diets.</caption><thead><tr><th rowspan="2"></th><th colspan="4">Glycerol Supplementation</th></tr><tr><th>0</th><th>2.5%</th><th>5.0%</th><th>7.5%</th></tr></thead><tbody><tr><td>Corn</td><td>57.67</td><td>54.82</td><td>51.97</td><td>49.14</td></tr><tr><td>Extracted rice bran</td><td>6.0</td><td>6.0</td><td>6.0</td><td>6.0</td></tr><tr><td>Soybean oil</td><td>1.05</td><td>0.95</td><td>0.85</td><td>0.75</td></tr><tr><td>SBM 49%</td><td>2.46</td><td>2.51</td><td>2.55</td><td>2.59</td></tr><tr><td>Glycerol</td><td>0</td><td>2.5</td><td>5.0</td><td>7.5</td></tr><tr><td>L-lysine</td><td>0.09</td><td>0.09</td><td>0.09</td><td>0.09</td></tr><tr><td>DL-methionine</td><td>0.145</td><td>0.145</td><td>0.145</td><td>0.145</td></tr><tr><td>Choline chloride 50%</td><td>0.007</td><td>0.007</td><td>0.007</td><td>0.007</td></tr><tr><td>Monodical P21</td><td>1.466</td><td>1.466</td><td>1.466</td><td>1.466</td></tr><tr><td>CaCO<sub>3</sub></td><td>8.36</td><td>8.36</td><td>8.36</td><td>8.36</td></tr><tr><td>Salt</td><td>0.36</td><td>0.36</td><td>0.36</td><td>0.36</td></tr><tr><td>Px</td><td>0.25</td><td>0.25</td><td>0.25</td><td>0.25</td></tr><tr><td>Chemical composition (analyzed)</td><td></td><td></td><td></td><td></td></tr><tr><td>GE, kcal</td><td>4023.79</td><td>3976.99</td><td>3995.37</td><td>3980.73</td></tr><tr><td>Crude Protein, %</td><td>17.41</td><td>17.65</td><td>17.48</td><td>17.51</td></tr><tr><td>Calcium, %</td><td>3.15</td><td>3.32</td><td>3.26</td><td>3.17</td></tr><tr><td>Phosphorus, %</td><td>0.66</td><td>0.63</td><td>0.61</td><td>0.60</td></tr></tbody></table>

**Results**

During the experimental period % mortality was not affected by treatment. These data are consistent with the study of CERRATE *et al.* (2006). In this study hens fed diet with purified glycerol at 2.5, 5.0 and 7.5% had no significantly affected on feed intake, hen house production, hen day production and egg weight in each period (Table 2). These data are consistent with the study of YALCIN *et al.* (2010) who found that feeding glycerol at 2.5-- 7.5% did not significantly affect body weight, egg production, egg weight and feed efficiency of laying hens. Similarly, CERRATE et al. (2006) found that the inclusion of 2.5 or 5.0% glycerol had no significant adverse effects on feed conversion at any age when compared to control diet (0% glycerol). However, using 10% glycerol in diet caused a reduction the body weight gain.

**Conclusion**

The inclusion of refined glycerol from biodiesel production up to 7.5% in diets had no adverse effects on feed intake and egg production. Therefore, purified glycerol can be effectively used as an energy source in layer diet.

**Acknowledgement**

The authors expressed their appreciation to Agricultural Research Development Agency (Public Organization) for financial support.
<table><caption>**Table 2.** The effects of dietary supplementation of glycerol on feed intake and egg production.</caption><thead><tr><th rowspan="2"></th><th colspan="4">Glycerol Supplementation (%)</th><th rowspan="2">P-value</th><th rowspan="2">Pooled SE</th></tr><tr><th>0</th><th>2.5</th><th>5.0</th><th>7.5</th></tr></thead><tbody><tr><td colspan="7"><strong>Period 1</strong></td></tr><tr><td>FI</td><td>108.95</td><td>105.82</td><td>106.05</td><td>109.36</td><td>0.327</td><td>0.3681</td></tr><tr><td>Hen day production (%)</td><td>92.76</td><td>91.91</td><td>93.19</td><td>93.39</td><td>0.9001</td><td>0.2139</td></tr><tr><td>Hen house production (%)</td><td>92.56</td><td>91.91</td><td>93.19</td><td>93.39</td><td>0.8910</td><td>0.2853</td></tr><tr><td>Egg weight (g)</td><td>61.72</td><td>60.94</td><td>61.38</td><td>61.15</td><td>0.1287</td><td>0.1364</td></tr><tr><td colspan="7"><strong>Period 2</strong></td></tr><tr><td>FI</td><td>112.72</td><td>113.63</td><td>112.19</td><td>114.98</td><td>0.6255</td><td>0.3545</td></tr><tr><td>Hen day production (%)</td><td>86.37</td><td>85.83</td><td>88.11</td><td>88.01</td><td>0.7234</td><td>0.2606</td></tr><tr><td>Hen house production (%)</td><td>83.45</td><td>84.23</td><td>83.16</td><td>86.76</td><td>0.7139</td><td>0.3378</td></tr><tr><td>Egg weight (g)</td><td>61.20</td><td>62.73</td><td>61.15</td><td>62.24</td><td>0.0813</td><td>0.4279</td></tr><tr><td colspan="7"><strong>Period 3</strong></td></tr><tr><td>FI</td><td>115.32</td><td>115.48</td><td>117.08</td><td>118.60</td><td>0.6326</td><td>0.4001</td></tr><tr><td>Hen day production (%)</td><td>86.97</td><td>89.97</td><td>91.02</td><td>89.09</td><td>0.5221</td><td>0.3265</td></tr><tr><td>Hen house production (%)</td><td>83.60</td><td>87.11</td><td>86.58</td><td>87.74</td><td>0.8015</td><td>0.2150</td></tr><tr><td>Egg weight (g)</td><td>61.08</td><td>64.97</td><td>63.64</td><td>63.94</td><td>0.2303</td><td>0.2644</td></tr><tr><td colspan="7"><strong>Period 4</strong></td></tr><tr><td>FI</td><td>104.03</td><td>104.61</td><td>103.07</td><td>103.73</td><td>0.9782</td><td>0.4520</td></tr><tr><td>Hen day production (%)</td><td>84.27</td><td>85.59</td><td>85.07</td><td>83.51</td><td>0.9175</td><td>0.3279</td></tr><tr><td>Hen house production (%)</td><td>83.96</td><td>81.34</td><td>77.47</td><td>83.17</td><td>0.4608</td><td>0.2554</td></tr><tr><td>Egg weight (g)</td><td>62.62</td><td>63.98</td><td>63.41</td><td>60.78</td><td>0.3585</td><td>0.2833</td></tr></tbody></table>

## Literature

Anonymous. 2007. Biodiesel Production. Thongma Biodiesel, Biodiesel from used cooking oil. http://thongmabiodiesel.blogspot.com/2007/05/naoh-0.html, 1 August, 2008.

CERRATE, S., F. YAN, Z. WANG, C. COTO, P. SACAKLI and P.W. WALDGROUP. 2006. Evaluating of glycerin from biodiesel production a feed ingredient for broilers. International Journal of Poultry Science.11:1001-1007.

HAYYAN, M., S.M. FAROUQ, A.H. MOHD and M.A. INAS. 2010. A novel technique for separating glycerine from palm-oil based biodiesel using ionic liquids. Elsivier. 91:116-120.

MA. F., M.A. HANNA. 1999. Biodiesel Production a review. Bioresource Technology. Elsivier. 70:1-5.

YALCIN, S., H. EROL, B. Özsoy, İ. Onbaşılar, S. Yalçın, A. Üner. 2010. Effects of glycerol on performance, egg traits, some blood parameters and antibody production to SRBC of laying hens. Elsivier. 129:129-134.
P-162 (ID 201)

EFFECT OF DIFFERENT NSP-DEGRADING ENZYME PRODUCTS IN TWO DIET TYPES ON GUT PARAMETERS AND BROILER PERFORMANCE

R.A.H.M. ten Doeschate<fnref n="1" /> and K. van de Belt<fnref n="2" />

**Corresponding author:** rob.tendoeschate@abvista.com

**Abstract**

The effects of five non-starch polysaccharide (NSP)-degrading enzyme products (differing in claimed activities but all containing xylanase as the main activity) against a control (no enzyme) were studied in two different diet types, one wheat/soya based diet (simple) and one diet with a more varied ingredient mix (complex), also including peas, distillers dried grains with solubles, sunflower meal and rapeseed. 2112 male Ross 308 day old broiler chicks were weighed and assigned to one of 8 replicates for each of the 12 treatments (22 per floor pen of 2.2 m<fnref n="2" />). Diets were iso-nutrient, contained phytase and coccidiostats as standard but no growth promoters. As enzyme addition may affect caecal fermentation, at 25 days of age the weight and length of the small intestine and ceaca were measured. The weight of the full intestine relative to the weight of the full caeca was reduced in the presence of the single xylanase enzymes (P=0.03), suggesting improved caecal fermentation. Growth rate was not affected by enzyme addition. FCR (0-42 days) showed a significant (P<0.001) diet*enzyme interaction, with only one of the five enzyme products showing an improved FCR in both diet types. There appeared to be no benefit from using a multi-activity product compared to the use of a single xylanase product in either diet, demonstrating that the benefits of NSP-enzyme addition can be achieved from xylanase alone.

**Keywords:** xylanase, broiler, alternative ingredients

**Introduction**

NSP-degrading enzymes are commonly used in a variety of broiler diets across the world. Most of the commercially available products contain xylanase as a major activity, but it is often argued that a complex diet would require an enzyme complex to improve performance rather than just a xylanase. Understanding the mode of action by which NSP enzymes improve nutrient digestibility and performance may assist the debate on whether a single xylanase product works across diet types or whether a wide spectrum of activities is needed to achieve this. Reduction of intestinal viscosity has long been considered a main target for NSP enzymes, and this is relevant in high viscous diets. Xylanase also might increase digesta retention time through the breakdown products from arabinoxylans effecting changes in the caecal microbiome and subsequently inducing a hormonal effect on gastric retention time (Singh et al., 2012). This mechanism would not require an enzyme complex to achieve improved performance across a range of diets, but suggests a single xylanase could have the same effect. The current study was set up to compare five commercial NSP enzyme products (differing in claimed activities but all containing xylanase as the main activity) against a control (no enzyme) in two different diet types, one wheat/soya based diet (simple) and one diet with a more varied ingredient mix (complex), also including peas, distillers dried grains with solubles, sunflower meal and rapeseed.

**Materials and Methods**

2112 male Ross 308 day old broiler chicks were weighed and assigned to one of 8 replicates for each of the 12 treatments (22 per floor pen of 2.2 m<fnref n="2" />). The birds were fed a starter diet from day 0-14, a grower diet from day 14-28 and a finisher diet from day 28-42. The two diet types were iso-nutrient, contained phytase and coccidiostats (Nicarban+Narasin in starter and Narasin in grower and finisher) as standard but no growth promoters. Basal diet composition is shown in table 1. The enzyme treatments were based on commercial products available in the European market, defined as follows: Control (no NSP enzyme); Xyl E (single xylanase product); Multi R (multi-enzyme product), Xyl H (single xylanase product), Xyl + β-Glu A (blend of xylanase and β-glucanase), Xyl W (single xylanase product). Each product was included at recommended levels for wheat based diets.

<footnote n="1">¹AB Vista, 3 Woodstock Court, Blenheim Road, Marlborough SN8 4AN, UK</footnote>

<footnote n="2">² Schothorst Feed Research, Lelystad, the Netherlands</footnote>
Birds and feed were weighed at each feed transition, on day 35 and at the end of the experiment at day 42. At 27 days of age two birds per pen were euthanized; weight and length of the small intestine and ceaca were measured and viscosity of jejunal digesta was measured.

<table><caption>**Table 1: Basal diet composition (%)**</caption><thead><tr><th rowspan="2"></th><th colspan="2">Starter<br>(0-14 days)</th><th colspan="2">Grower<br>(14-28 days)</th><th colspan="2">Finisher<br>(28-42 days)</th></tr><tr><th>Simple</th><th>Complex</th><th>Simple</th><th>Complex</th><th>Simple</th><th>Complex</th></tr></thead><tbody><tr><td>Wheat</td><td>59.2</td><td>48.1</td><td>64.0</td><td>50.8</td><td>67.6</td><td>54.5</td></tr><tr><td>Soybean meal</td><td>30.3</td><td>23.2</td><td>27.8</td><td>17.2</td><td>23.9</td><td>12.8</td></tr><tr><td>Maize DDGS</td><td>-</td><td>5.0</td><td>-</td><td>5.0</td><td>-</td><td>5.0</td></tr><tr><td>Peas</td><td>-</td><td>5.0</td><td>-</td><td>5.0</td><td>-</td><td>5.0</td></tr><tr><td>Fish meal</td><td>2.5</td><td>2.5</td><td>-</td><td>-</td><td>-</td><td>-</td></tr><tr><td>Sunflower meal</td><td>-</td><td>4.0</td><td>-</td><td>7.5</td><td>-</td><td>7.5</td></tr><tr><td>Rape seed</td><td>-</td><td>5.0</td><td>-</td><td>7.5</td><td>-</td><td>8</td></tr><tr><td>Soybean oil</td><td>1.1</td><td>1.0</td><td>1.0</td><td>1.0</td><td>1.8</td><td>1.0</td></tr><tr><td>Animal Fat</td><td>2.1</td><td>1.6</td><td>3.3</td><td>2.3</td><td>2.3</td><td>1.9</td></tr><tr><td>Palm oil</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1.0</td><td>1.0</td></tr><tr><td>Monocalcium phosphate</td><td>0.59</td><td>0.48</td><td>0.39</td><td>0.26</td><td>0.22</td><td>0.08</td></tr><tr><td>Salt</td><td>0.19</td><td>0.15</td><td>0.22</td><td>0.15</td><td>0.19</td><td>0.09</td></tr><tr><td>Limestone</td><td>1.49</td><td>1.49</td><td>1.25</td><td>1.23</td><td>1.09</td><td>1.06</td></tr><tr><td>Sodium bicarbonate</td><td>0.03</td><td>-</td><td>-</td><td>-</td><td>0.06</td><td>0.11</td></tr><tr><td>Premix complex<sup>1</sup></td><td>2.43</td><td>2.47</td><td>1.95</td><td>2.06</td><td>1.80</td><td>1.94</td></tr><tr><td colspan="7">Calculated nutrients</td></tr><tr><td>Crude Protein (g/kg)</td><td>241</td><td>243</td><td>217</td><td>219</td><td>202</td><td>202</td></tr><tr><td>AME broiler (Kcal/kg)</td><td>2800</td><td>2800</td><td>2900</td><td>2900</td><td>3000</td><td>3000</td></tr><tr><td>Digestible Lysine (g/kg)</td><td>12.5</td><td>12.5</td><td>11.2</td><td>11.2</td><td>10.0</td><td>10.0</td></tr><tr><td>Digestible M+C (g/kg)</td><td>9.3</td><td>9.3</td><td>8.5</td><td>8.5</td><td>7.8</td><td>7.8</td></tr><tr><td>Digestible Thr (g/kg)</td><td>8.1</td><td>8.1</td><td>7.1</td><td>7.1</td><td>6.4</td><td>6.4</td></tr><tr><td>Calcium (g/kg)</td><td>8.4</td><td>8.4</td><td>6.4</td><td>6.4</td><td>5.4</td><td>5.4</td></tr><tr><td>Ret P<sup>2</sup> (g/kg)</td><td>4.0</td><td>4.0</td><td>3.2</td><td>3.2</td><td>2.8</td><td>2.8</td></tr></tbody></table>

## Results and Conclusion

Nutrient and enzyme levels in the diets were within acceptable limits of variation. Intestinal viscosity was not affected by diet or enzyme (table 2), which means enzyme benefits can not be explained by a reduction in digesta viscosity.

At 42 days of age (table 2) there was a significant (P<0.001) interaction between diet type and enzyme product for FCR. One single xylanase product gave a significantly improved FCR in both diet types (simple: 1.600 cf 1.642; complex: 1.618 cf 1.643) whilst in the simple diet three of the products showed no significant improvement and in the complex diet one of the other products showed no significant improvement.

The weight of the full small intestine relative to the weight of the full caecum was significantly reduced by the three single xylanase enzyme products as compared to the control diet. A reduction in this ratio can indicate a more active caecal fermentation, which could be the result of the supply of arabinoxylan-oligosaccharides, as discussed by Singh et al (2012). This finding thus supports the hypothesis that a single xylanase product can work across a variety of diet types.

<footnote n="1">1) Premix complex comprises vitamins, trace minerals, amino acids, coccidiostats and phytase.</footnote>

<footnote n="2">2) Calculated retainable P includes contribution from phytase</footnote>
<table><caption>Table 2: Results</caption><thead><tr><th colspan="2"></th><th>Gain 0-42 d (g)</th><th>FCR 0-42 d (g:g)</th><th>Digesta Viscosity (cPs)</th><th>Intestine Wt relative to caecum</th></tr></thead><tbody><tr><th colspan="6" style="text-align: center;">Interaction Effect</th></tr><tr><th>Diet</th><th>Enzyme</th><th></th><th></th><th></th><th></th></tr><tr><td>Simple</td><td>Control</td><td>3379</td><td>1.642<sup>cd</sup></td><td>2.84</td><td>9.0</td></tr><tr><td>Simple</td><td>Xyl E</td><td>3413</td><td>1.600<sup>a</sup></td><td>2.60</td><td>7.1</td></tr><tr><td>Simple</td><td>Multi R</td><td>3387</td><td>1.602<sup>a</sup></td><td>2.63</td><td>8.1</td></tr><tr><td>Simple</td><td>Xyl H</td><td>3387</td><td>1.640<sup>bcd</sup></td><td>3.11</td><td>8.8</td></tr><tr><td>Simple</td><td>Xyl + β-Glu A</td><td>3440</td><td>1.619<sup>abc</sup></td><td>2.71</td><td>8.5</td></tr><tr><td>Simple</td><td>Xyl R</td><td>3379</td><td>1.646<sup>d</sup></td><td>2.94</td><td>7.4</td></tr><tr><td>Complex</td><td>Control</td><td>3366</td><td>1.643<sup>d</sup></td><td>2.60</td><td>9.6</td></tr><tr><td>Complex</td><td>Xyl E</td><td>3322</td><td>1.618<sup>ab</sup></td><td>2.83</td><td>8.1</td></tr><tr><td>Complex</td><td>Multi R</td><td>3319</td><td>1.648<sup>d</sup></td><td>3.15</td><td>9.2</td></tr><tr><td>Complex</td><td>Xyl H</td><td>3364</td><td>1.619<sup>ab</sup></td><td>2.76</td><td>7.5</td></tr><tr><td>Complex</td><td>Xyl + β-Glu A</td><td>3334</td><td>1.619<sup>ab</sup></td><td>2.79</td><td>9.0</td></tr><tr><td>Complex</td><td>Xyl R</td><td>3298</td><td>1.612<sup>a</sup></td><td>2.83</td><td>7.1</td></tr><tr><td></td><td>LSD</td><td>74.8</td><td>0.023</td><td>0.765</td><td>1.59</td></tr><tr><td></td><td>P</td><td>0.421</td><td>&lt;0.001</td><td>0.620</td><td>0.255</td></tr><tr><th colspan="6" style="text-align: center;">Diet Effect</th></tr><tr><td>Simple</td><td></td><td>3397<sup>b</sup></td><td>1.625</td><td>2.80</td><td>8.2</td></tr><tr><td>Complex</td><td></td><td>3334<sup>a</sup></td><td>1.626</td><td>2.83</td><td>8.4</td></tr><tr><td></td><td>LSD</td><td>30.5</td><td>0.0094</td><td>0.765</td><td>0.65</td></tr><tr><td></td><td>P</td><td>&lt;0.001</td><td>0.725</td><td>0.620</td><td>0.407</td></tr><tr><th colspan="6" style="text-align: center;">Enzyme effect</th></tr><tr><td>Control</td><td></td><td>3372</td><td>1.642</td><td>2.72</td><td>9.3<sup>d</sup></td></tr><tr><td>Xyl E</td><td></td><td>3368</td><td>1.609</td><td>2.71</td><td>7.6<sup>ab</sup></td></tr><tr><td>Multi R</td><td></td><td>3353</td><td>1.625</td><td>2.89</td><td>8.6<sup>bcd</sup></td></tr><tr><td>Xyl H</td><td></td><td>3375</td><td>1.629</td><td>2.94</td><td>8.2<sup>abc</sup></td></tr><tr><td>Xyl + β-Glu A</td><td></td><td>3387</td><td>1.619</td><td>2.75</td><td>8.7<sup>cd</sup></td></tr><tr><td>Xyl R</td><td></td><td>3338</td><td>1.629</td><td>2.88</td><td>7.2<sup>a</sup></td></tr><tr><td></td><td>LSD</td><td>52.9</td><td>0.0163</td><td>0.541</td><td>1.12</td></tr><tr><td></td><td>P</td><td>0.511</td><td>0.004</td><td>0.931</td><td>0.003</td></tr></tbody></table>

References

SINGH, A., H.V. MASEY O'NEILL, T.K. GHOSH, M.R. BEDFORD and S. HALDAR, 2012: Effect of xylanase supplementation on performance, total volatile fatty acids and selected bacterial population in caeca, metabolic indices and peptide YY concentrations in serum of broiler chickens fed energy restricted maize-soybean based diets. Animal Feed Science and Technology 177, 194-203
# Mineral nutrition

P-166 (ID 309)

## DIETARY SELENIUM SUPPLEMENTATION OF BREEDERS INFLUENCES GROWTH AND SELENIUM STATUS OF PROGENY

M. Briens¹, F. Couloigner¹, F. Rouffineau¹, P.A. Geraert² and Y. Mercier¹

¹Adisseo France S.A.S., CERN, 6, route noire, Malicorne, France

² Adisseo France S.A.S., 10, Place du Général de Gaulle, 92160 Antony, France

**Corresponding author:** yves.mercier@adisseo.com

### Abstract

The impact of selenium (Se) sources fed to breeders on muscle Se status and progeny growth was studied. Breeders were divided into 3 groups differing by the dietary Se source as followed: sodium selenite (SS), selenized-yeast (SY) and hydroxy-selenomethionine or HMSeBA (SO). Each breeder group generated 290 day-old chick. At d0, 30 chicks per group were selected to measure muscle Se content and the 260 remaining chicks were divided into 13 replicate pens of 20 chicks for 21 days. Progeny of the 3 groups all received the same standard broilers diet containing only SS. On D12 and 21, growth performances were measured and one chick per pen was selected to measure breast Se content. Breeder Se source had no effect on the body weight at hatch, body weight gain or feed intake during the entire experimental period (P > 0.05). However, feed conversion ratio (FCR) of progeny issued from SO fed breeders was significantly improved between D0 to 12 and D0 to 21 comparatively to SS (P < 0.05). Muscle Se content of day-old broiler chicks was higher for SO group compared to other treatments (P < 0.05). In addition, muscle Se content of day-old broiler appeared to be significantly related to FCR for the periods: D0 to 12 and D0 to 21 (P < 0.01). These results showed the higher ability of HMSeBA compared to other Se source in breeder diet to improve chick Se status at hatch which is related to FCR improvement during 0-21 day period of growth.

**Keywords:** selenium, hydroxy-selenomethionine, broiler breeder, progeny

### Introduction

Several studies showed the benefits of dietary organic selenium (Se) supplementation in breeders on egg quality and fertility compared to inorganic Se (Pappas et al. 2005a). However, few studies observed the effect of breeder nutrition on the performance of the offspring. Indeed, young broiler chicks are facing strong oxidative stress during their first days of life due to important physiological changes, such as, adaptation to aerobic life, active development of physiological systems such as digestive tract, hormonal and neuronal systems (Moran 2007). Surai (2000) had shown an increase of the Se content of day-old-chick at hatch and higher glutathione level and glutathione peroxidase (GPx) activity in liver of their chicks at least 5 days after hatch when breeders were supplemented with selenized yeasts. Moreover, Pappas et al. (2005b) had observed a positive effect of the breeders Se intake on the GPx activity in tissues of their offspring throughout the 4 weeks following hatch. Recently, Jlali et al. (2013) demonstrated that egg selenium content can be tremendously increased with different organic selenium sources when compared to inorganic selenium sources, resulting in selenium deposition higher with hydroxy-selenomethionine (HMSeBA) compared to selenized yeast. The present work aimed evaluating the effect of breeder selenium sources on selenium status and growth performance of their progeny.

### Materials and Methods

A total of 870 day-old chicks from 3 different broiler breeders groups (3 x 290) were used in this study. The 3 breeder groups differing in dietary Se sources and level as followed: sodium selenite at 0.3 mg Se/kg (SS); selenized yeast at 0.2 mg Se/kg (SY); hydroxy-selenomethionine or HMSeBA at 0.2 mg Se/kg (SO). On the 290 day-old chicks per group, 30 were randomly selected and euthanized on the arrival of birds to measure their birth Se status. The pectoralis major were collected and pooled by 5 chicks in order obtaining 6 pools of muscle per breeder groups. The 260 remaining chicks were allocated in 13 pen replicates of 20 chicks for 21 days. Whatever the origin of the chicks, they received the same starter and grower commercial broiler diets containing only sodium selenite as Se supplementation at 0.2 mg Se/kg of feed. The body weight, feed intake and FCR of birds were measured on day 12 and 21 and muscle samples have been collected on 1 bird per pen at D12 and D21 to follow Se status of birds. The total selenium concentration in
diets and muscles was determined by inductively coupled mass spectrometry (ICP-MS) according the method of Bierla et al. (2008). All data were analyzed using GLM procedure of SAS 9.1.3 (SAS Inst. Inc., Cary, NC).

## Results

The measurement of Se level of the breeder diets confirmed targeted levels (data not shown). The progeny starter and grower broiler diets naturally contained 0.08 and 0.11 mg Se/kg, respectively and resulted to 0.26 and 0.35 mg Se/kg, respectively after SS supplementation.

<table><caption>**Table 3:** Broiler growth performance in relation with selenium sources distributed to breeders</caption><thead><tr><th>Breeders diet</th><th>SS</th><th>SY</th><th>SO</th><th>SEM</th><th>P-value</th></tr></thead><tbody><tr><td>D0-12</td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>BW D0 (g)</td><td>40.31</td><td>40.23</td><td>40.31</td><td>0.19</td><td>0.876</td></tr><tr><td>BW D12 (g)</td><td>327.4</td><td>328.3</td><td>331.0</td><td>43.7</td><td>0.366</td></tr><tr><td>BWG (g)</td><td>287.1</td><td>288.1</td><td>290.7</td><td>45.0</td><td>0.381</td></tr><tr><td>FI (g)</td><td>357.4</td><td>354.0</td><td>355.9</td><td>60.6</td><td>0.544</td></tr><tr><td>FCR (g/g)</td><td>1.245<sup>a</sup></td><td>1.229<sup>ab</sup></td><td>1.225<sup>b</sup></td><td>0.0003</td><td>0.025</td></tr><tr><td>D12-21</td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>BW D21 (g)</td><td>710.6</td><td>711.6</td><td>717.5</td><td>180.7</td><td>0.389</td></tr><tr><td>BWG (g)</td><td>383.2</td><td>383.3</td><td>386.5</td><td>75.9</td><td>0.566</td></tr><tr><td>FI (g)</td><td>613.6</td><td>610.4</td><td>612.8</td><td>124.7</td><td>0.743</td></tr><tr><td>FCR (g/g)</td><td>1.602</td><td>1.593</td><td>1.586</td><td>0.0003</td><td>0.096</td></tr><tr><td>D0-21</td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>BWG (g)</td><td>670.3</td><td>671.4</td><td>677.2</td><td>182.3</td><td>0.395</td></tr><tr><td>FI (g)</td><td>971.0</td><td>964.4</td><td>968.7</td><td>277.0</td><td>0.591</td></tr><tr><td>FCR (g)</td><td>1.449<sup>a</sup></td><td>1.436<sup>ab</sup></td><td>1.431<sup>b</sup></td><td>0.0002</td><td>0.007</td></tr></tbody></table>

a, b: Means with different superscripts are statically different (P < 0.05)

The growth performances of day-old chicks are presented in Table 1. The breeder Se sources had no effect on hatch weight of the offspring, feed intake (FI), body weight gain (BWG). However, broilers from breeders fed with SO had a better FCR between D0 to D12 and D0 to D21 (P < 0.05) compared to the broilers from breeders fed with SS, while chicks issued from SY treatment presented intermediary FCR. Chick muscle Se content at the different sampling days are presented in Figure 1A. At hatch, broilers from breeders fed with organic Se source (SO and SY) had a higher muscle Se content (P < 0.001) compared to SS maternal treatment, with an additional concentration using SO compared to SY treatment (P < 0.001). These results are consistent with those of Jlali et al. (2013) who reported egg Se content in the same hierarchy with those sources. The link between egg selenium content and day old chick selenium status had been also reported by Surai (2000). However, after 12 days, muscle Se content was no longer discriminating maternal treatments. The significant decrease from D0 to D12 could be explained, on one hand by the low efficiency of inorganic Se source supplied during growth to maintain muscle Se content, and, on the other hand by the selenium turnover to build selenoprotein, such as GPx, for antioxidant defenses (Surai, 2002). Our results corroborates a study of Wang et al. (2011) who fed sodium selenite or selenomethionine in breeder and studied the offspring performances. They also reported significantly higher antioxidant status of chicks from organic selenium group with higher GPx and SOD activities in breast muscle compared to inorganic group.
A

<figure><img src="image_63.png" /></figure>

B

<figure><img src="image_64.png" /><figcaption>**Figure 3:** A) Evolution of the broilers muscle Se content in relation with dietary selenium source of breeders.</figcaption></figure>

B) Effect of muscle selenium content at hatch on 0-21 days FCR in broilers

SO significantly improved FCR during the 0-21 day period compared to SS. The three different selenium sources supplied to breeder were related to progeny breast muscle selenium concentration and FCR (Figure 1B). Day old chick Se content had a significant effect on the FCR of broilers between D0 to D12 and between D0 to D21 (P < 0.01). The improvement of broilers FCR with the use of organic selenium forms in breeder feeds could be explained by higher muscle Se reserves that allowed birds to manage selenium demand during the early growth phase and ensure better seLENOProtein production to maintain their antioxidant system (Pappas et al. 2005b, Wang et al 2011). These results point out the importance of day old chick selenium status on their subsequent performances. This resulted in an average of FCR improvement of 1.25 % during the 0-21 day period for broilers from SO treatment compared to broilers from SS treatment.

## Conclusions

This study showed the higher ability of HMSeBA to supply selenium through maternal diet into eggs and chick progeny comparatively to seleno-yeasts and sodium selenite. Day old chick selenium status was correlated to 0-21 day FCR, highlighting the importance of selenium status for optimal performances and certainly maintain antioxidant defenses during this critical phase of chick early life.

## Literature

BIERLA, K., DERNOVICS, M., VACCHINA, V., SZPUNAR, J., BERTIN, G., AND LOBINSKI, R., 2008: *Analytical Bioanalytical Chemistry*. **390**: 1789–1798.

JLALI M., BRIENS M., ROUFFINEAU F., MERCERAND F., GERAERT P.-A. AND MERCIER Y., 2013: *Journal of Animal Science*. **91**: 1745-1752.

MORAN E.T., 2007: *Poultry Science*. **5**:1043-1049.

PAPPAS A.C., ACAMOVIC T., SPARKS N.H., SURAI P.F. AND MCDEVITT R.M., 2005a: *Poultry Science*. **84**: 865-874.

PAPPAS A.C., KARADAS F., SURAI P.F. AND SPEAKE B.K., 2005b: *Comparative Biochemistry Physiology- Part B Biochemistry & Molecular Biology*. **142**: 465-474.

SURAI P.F., 2000: *British Poultry Science*. **41**:235-243.

WANG, Y.X., ZHAN, X.A., YUAN, D., ZHANG, X.W., WU R.J., 2011: Biological Trace Element. Research. **143**: 1497-1507.

YOON, I., T. M. WERNER, AND J. M. BUTLER., 2007: *Poultry Science*. **86**:727-730.
P-167 (ID 308)

TOWARD AN EASY ASSESSING METHOD TO MEASURE SELENIUM BIOAVAILABILITY IN BIRDS

M. Briens<fnref n="1" />, F. Couloigner<fnref n="1" />, M. Jlali<fnref n="1" />, F. Rouffineau<fnref n="1" />, P.A. Geraert<fnref n="2" /> and Y. Mercier<fnref n="1" />

**Corresponding author:** yves.mercier@adisseo.com

**Abstract**

The aim of this study was to develop an easy assessment method to determine the selenium (Se) bioavailability of different selenium sources in birds. The first step was to measure the kinetic of selenium deposition in muscle and feathers of broilers fed different selenium sources. A total of 576 chicks were divided into 4 treatments with 8 replicates of 18 birds per pen for 21 days. The diets used in the experiment were supplemented with different Se sources as followed: negative control (NC) not supplemented in Se; sodium selenite (SS); selenized yeast (SY) and hydroxy-selenomethionine or HMSeBA (SO) at 0.2 mg Se/kg of feed. Total selenium content in breast muscle and feathers was assessed on day 0, 7, 14 and 21. Only SO increased muscle Se content at D7 compared to D0 (P < 0.05), while all other treatments exhibited decrease of Se muscle content (P < 0.05) from D0 to D7. At D21, the muscle and feather Se content were highly correlated (R<fnref n="2" /> = 0.927; P < 0.0001). This study shows that breast muscle selenium measured as early as at D7 allows discriminating selenium sources as well as at D21. Moreover, the feather Se content measured at 21 days appeared a good and convenient indicator of selenium status in birds that can be used at farm level. These results also underline the benefit of a pure organic selenium source such as hydroxy-selenomethionine to rapidly increase selenium status in early growth phase.

**Keywords:** selenium, hydroxy-selenomethionine, feather, poultry

**Introduction**

The tissues of chick embryos and newly hatched chicks contain a high proportion of polyunsaturated fatty acids. During their first days of life, birds face a rather stressful period, and are highly susceptible to oxidative stress (Surai and Fisinin, 2014). In addition, their hepatic concentration of vitamin E and carotenoids decrease by 20-fold during the 10 first days of life (Surai 2002). However, the glutathione peroxidase (GPx) liver activity increased in the same time. In practice, signs of selenium (Se) deficiency are generally observed around 7 days post-hatch, when vitamin E level is low and GPx activity not yet sufficient (Surai, 2002). Traditionally, to ensure adequate antioxidant status of young broilers, Se supplementation is provided in the diets, using sodium selenite (SS) or organic forms like seleno-yeasts (SY) containing mainly SeMet in variable amount according to the strain and growing yeast conditions (Surai and Fisinin, 2014; Simon et al. 2013). The Se status of wild birds is currently evaluated with the measurement of Se content of their feathers (Burger, 1996). The purpose of this study was to evaluate the possibility to use muscle or feather Se contents to compare the bioavailability of different selenium sources.

**Material and Methods**

A total of 576 day-old chicks from a commercial hatchery were allocated to 4 treatments with 8 replicates of 18 birds per pen. The birds received a starter diet from D0 to D11 and a grower diet from D12 to D21 and each treatment was supplemented with different Se sources as follow: NC, basal diet not supplemented with Se; SS, SY and SO, basal diet supplemented with 0.2 mg Se/kg from sodium selenite, selenized yeast and HMSeBA respectively. At the end of each phase: body weight (BW) and pen feed intake (FI) were recorded and used for calculation of feed conversion ratio (FCR). Moreover, on D0, 7, 14 and 21, one bird per pen (8 per treatment) was randomly selected and euthanized. The feathers of the wings and the two *Pectoralis major* muscle were collected and pooled for two birds before being frozen at -20°C for subsequent analysis. The total Se concentration of diets, feathers and muscles were determined by inducively coupled mass spectrometry (ICP-MS) according to Bierla et al. (2008). All data were analyzed using GLM procedure of SAS 9.1.3 (SAS Inst. Inc., Cary, NC).

<footnote n="1">¹Adisseo France S.A.S., CERN, 6, route noire, Malicorne, France</footnote>

<footnote n="2">² Adisseo France S.A.S., 10, Place du Général de Gaulle, 92160 Antony, France</footnote>
## Results

The feed analyses indicated an expected native Se content of 0.13 and 0.10 mg Se/kg in starter and grower diet, respectively (data not shown). The different treatments had no effect on growth performance during the 21 days of experiment (average BW = 918 g and average FCR = 1.39 g/g). These results are consistent with other works that did not show effects on performance in relation with dietary selenium (Se) supplementation in broilers under standard conditions (Briens et al. 2013).

The evolution of muscle Se content through the experiment depending on the Se source is presented in Figure 1. Selenium sources significantly affected the Se deposition in *pectoralis major* muscle (P < 0.001) at D7 and the same effects remained until 21 days old. At D7, chicks fed with the diet supplemented with SO treatment showed a significant increase of muscle Se content compared to hatch level (+ 41%) whereas other treatments showed significant decrease in muscle selenium content (-15, -37 and -43 % for SY, SS and NC respectively). SY supplementation restored muscle selenium content after 14 days at a similar level than D0. SS treatment appeared to keep muscle selenium content stable from D7 to D21, whereas for NC treatment muscle selenium content decreased until D14. These results are consistent with previous works studying muscle Se concentration with different Se sources supplemented for 21 day (Briens et al. 2013, 2014). From D7 to D21, the muscle Se content of broilers fed SS diet remains stable, whereas the muscle Se content of broilers fed NC continued to decrease. This finding showed that even a relatively high feed Se basal level (0.13 to 0.10 mg Se/kg in NC) cannot maintain Se status of broilers. The slight decrease of muscle Se content of chicks fed with SY during the first days of life and the subsequent recovery between D7 and D14 could be related to the Se form in yeasts, SeMet incorporated in proteins, that may be not optimally digested during the first week because of suboptimal gastrointestinal functions of the young chicks whereas a free seleno amino acid like SO may be directly absorbed.

<figure><img src="image_65.png" /><figcaption>**Figure 4:** Kinetic of muscle Se content in *Pectoralis major* muscle in function of the dietary Se source. Results are expressed as mean ± SD and different letters indicate statistical differences at P < 0.05.</figcaption></figure>

Se concentration in feathers collected on day 7 and 21 on birds fed NC, SY and SO diet was summarized in Figure 2A. Whatever the dietary treatment, the Se concentration in feathers decreased (P < 0.001) with age of the birds. At both sampling time (7 or 21 days) feather Se content appeared significantly affected (p<0.05) by dietary Se content and source as follow: NC < SY < SO. The present results are in agreement with the study of Sevcikova et al. (2006). The correlations between muscle and feather Se content at D7 and D21 were highly significant (P < 0.001) with a better accuracy at D21 (R² = 0.927) than at D7 (R²= 0.542) (Figure 2B). This could be explained because at latter age the second generation of feathers starts to grow and push out the down feather (Leeson and Walsh, 2004). These results allow to propose to measure feather Se content at 21 days of age as a simple and accurate indicator to follow the muscle Se status of birds and to compare bioavailability of selenium sources.
<figure><img src="image_66.png" /><figcaption>**Figure 5:** A: Se deposition in feathers in function of dietary selenium source at day 7 and 21. Results are expressed as mean ± SD and different letters indicate statistical differences at P < 0.05; B: Correlation between Se deposited in feathers and in *Pectoralis major* muscle.</figcaption></figure>

**Conclusions**

The selenium status and the efficacy of different selenium sources can be evaluated through a non-invasive procedure: the measure of feather Se content. Muscle and feather Se content are highly correlated at D21, and transfer Se efficiency is dependent of the Se source. In those conditions HMSeBA showed the best ability to increase bird Se deposition, an indicator of their Se status.

**Literature**

BIERLA, K., DERNOVICS, M., VACCHINA, V., SZPUNAR, J., BERTIN, G., AND LOBINSKI, R., 2008: *Analytical Bioanalytical Chemistry* **390**: 1789–1798.

BRIENS, M., Y. MERCIER, F. ROUFFINEAU, V. VACCHINA, AND P.A. GERAERT., 2013: *British Journal Nutrition* **110**: 617-624.

BRIENS M., MERCIER Y., ROUFFINEAU F., MERCERAND F. AND GERAERT P.A., 2014: *Poultry Science* **93**: 85-93.

BURGER J., 1996: *The Auk*. **113**: 399-407.

LEESON S. AND WALSH T., 2004: *World's Poultry Science Journal*. **60**: 42-51

SEVCIKOVA S., M. SKRIVAN, G. DLOUHA AND KOUCYK M., 2006: *Czech Journal Animal Science* **51**: 449-457.

SIMON E., N. BALLET, M. FRANCESCH AND BRUFAU J., 2013: *World's Poultry Science Journal*. **69**, Supplement: 1-5

SURAI P.F. AND FISININ V.I., 2014: *Animal Feed Science Technology* **191**: 1-15

SURAI P.F., 2002: *World's Poultry Science Journal*. **58**: 333-347

SUZUKI K.T., 2005: *Journal Health Science* **51**:107-144.
P-171 (ID 79)

BROILER PERFORMANCE AND PREVALENCE OF FOOTPAD LESIONS IN RESPONSE TO PHYTASE, CALCIUM AND PHOSPHORUS CONCENTRATIONS

E. Delezie<fnref n="1" />, K. Bierman<fnref n="2" />, L. Nollet<fnref n="2" />, S. Leleu<fnref n="1" />, L. Maertens<fnref n="1" />.

Corresponding author: evelyne.delezie@ilvo.vlaanderen.be

Abstract

Taking into account the concerns about the environment, nutrition and farm profits, animals have to be fed precisely to meet their phosphorus (P) requirements. The objective of the current trial was to provide efficacy data of a 6-phytase for use in broilers at different calcium (Ca) levels and Ca/P ratios. Phytase was introduced at different dosage levels (100g/tonne and 200g/tonne) into diets i) with a standard Ca and reduced P level (imbalanced diet); ii) with a reduced Ca and P level or iii) with a normal Ca and P level (balanced diets). The performance of the balanced treatments without phytase supplementation did not differ significantly. The balanced diets (with and without phytase supplementation) had a significantly lower WAFC1500 (FC to obtain a body weight of 1500g) compared to the imbalanced diets. Adding phytase to the low Ca-low P diet did further increased the performance and the obtained results were even better (not significantly) than those fed the standard Ca and P diet. A higher dosage of phytase combined with an imbalanced diet had a significant effect as the higher supplementation level could compensate for the shortcomings of the imbalanced diet. The highest prevalence of footpad lesions was seen at broilers fed the imbalanced diets. The overall results emphasise that dietary Ca and P concentrations can be reduced but only in a balanced way. If done in combination with phytase even higher performance results were obtained than when using standard dietary Ca and P levels.

Introduction

Due to the environmental pressure, but also from a nutritional and economical point of view, the phosphorus (P)-content
of a diet is an increasingly important issue and animals must be fed near their P requirements. However, it is very diffi-
cult to know the exact P requirements of broilers as there are interactions between eg. the calcium (Ca) level, P level
and phytase (L'ETOURNEAU-MONTMINY et al., 2010). Therefore, the objective of this trial was to provide efficacy
data of a 6-phytase for use in broiler diets at different Ca levels and Ca/P ratio's.

Materials and Methods

One day old, male broiler chicks (Ross 308) were used in this trial (Origin: Belgabroed Hatchery, Merksplas-Belgium). For the trial, 36 pens were used and male birds were housed with 35 per pen. The floor was covered with fresh wood shavings and no additional filling or cleaning of the floor was executed during the trial. Central water heating and infrared bulbs (1 per pen) provide optimal house temperature. The experiment consisted of 6 dietary treatments with each 6 replicates of 35 birds or 1260 birds in total. The first dietary treatment consisted of a corn-soybean diet which met all dietary recommendations for chickens from 1 to 39 days of age. For the second, fifth and sixth treatment Ca and P level were reduced during the entire period. For the third and fourth treatment only the P level was reduced during the entire period. All treatments were formulated to be isocaloric and isonitrogenous. For treatment 3 and 5 and for treatment 4 and 6 Optiphos 2500 Granular was supplemented at 250 (100 g/tonne) and 500 OTU (200g/tonne), respectively. A 3-phase feeding scheme was applied with periods of 13 days each.

Average pen weight was recorded at day old, 13, 21, 26 and 39 days of age. Feed intake was recorded for 1-13, 13-21, 21-26 and 26-39 days. Feed conversion, daily growth rate, bird-days and daily feed intake per bird will be calculated for 1-13, 13-21, 21-26, 26-39 days and also for 1-26 and 1-39 days. Dead animals were recorded during the entire period. Foot pad dermatitis of 8 birds per pen was determined. Daily mortality and cullings were recorded for each pen. Corrections for mortality calculating zootechnical performances were done using the number of 'animal days' (number of animals x days alive).

All zootechnical parameters were analysed by a linear mixed model with treatment as a fixed factor and block as a random factor (Statistica 11.0 program). Data that differ significantly have a P-value below 0.05. The scoring data were analysed using a proportional odds logistic regression model for ordinal responses (polr function in MASS package in R 3.0.1).

<footnote n="1">¹Institute for Agricultural and Fisheries Research (ILVO), Scheldeweg 68, 9090 Melle, Belgium</footnote>

<footnote n="2">²Huvepharma Uitbreidingstraat 80, 2600 Antwerp, Belgium</footnote>
## Results

Main performance results are presented in table 1. Lowest body weight was obtained if broilers were given the imbalanced diet with low phytase supplementation (Tr3). Until d21 of age, body weight of broilers fed Tr3 was significantly lower compared to broilers fed Tr4 to Tr6. From d26 onwards, the difference in body weight of the broilers fed the imbalanced diets (Tr3 and Tr4) was no longer present. But body weight of Tr3 fed broilers remained significantly lower compared to the supplemented NC groups (Tr5 and Tr6). No significant differences between the balanced diets without phytase supplementation (Tr1 and Tr2) were observed until slaughter age. If broilers were given the normal Ca, low P diet (Tr2), body weight at slaughter age decreased with 32.9 g compared to their normal Ca, normal P fed counterparts (Tr1). Supplementing phytase at the NC groups (Tr4 and 5) significantly increased the body weight at d26 and d39. The higher supplementation level of phytase did not result in a further significant increase of body weight. However, a higher supplementation level for the unbalanced diet (Tr4 vs Tr3) significantly increased body weight until d21. This increase was no longer significant at the 5% level after d21 of age.

<table><caption>**Table 1:** Effect of dietary treatments on body weight (BW) (g), feed intake (g/day) and feed conversion during the starter (d0-d13), grower (d13-d26) and finisher (d26-d39) period.</caption><thead><tr><th></th><th>Tr1</th><th>Tr2</th><th>Tr3</th><th>Tr4</th><th>Tr5</th><th>Tr6</th><th>P-value</th></tr></thead><tbody><tr><td><strong>BW (g)</strong></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>d13 (g)</td><td>303.8<sup>bc</sup></td><td>305.7<sup>abc</sup></td><td>294.6<sup>c</sup></td><td>318.3<sup>ab</sup></td><td>326.9<sup>a</sup></td><td>315.5<sup>ab</sup></td><td>0.002</td></tr><tr><td>d26 (g)</td><td>1100.8<sup>a</sup></td><td>1083.1<sup>bc</sup></td><td>1027.3<sup>c</sup></td><td>1108.9<sup>a</sup></td><td>1129.2<sup>a</sup></td><td>1160.7<sup>a</sup></td><td>0.0004</td></tr><tr><td>d39 (g)</td><td>2290.4<sup>a</sup></td><td>2257.5<sup>bc</sup></td><td>2214.8<sup>c</sup></td><td>2297.4<sup>a</sup></td><td>2363.1<sup>a</sup></td><td>2351.8<sup>a</sup></td><td>0.0008</td></tr><tr><td><strong>FI (g/day)</strong></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>d0-d13</td><td>27.7<sup>b</sup></td><td>28.4<sup>ab</sup></td><td>28.4<sup>ab</sup></td><td>28.9<sup>ab</sup></td><td>30.2<sup>a</sup></td><td>28.9<sup>ab</sup></td><td>0.02</td></tr><tr><td>d13-d26</td><td>89.6<sup>ab</sup></td><td>88.0<sup>ab</sup></td><td>85.0<sup>b</sup></td><td>91.4<sup>a</sup></td><td>91.6<sup>a</sup></td><td>93.1<sup>a</sup></td><td>0.006</td></tr><tr><td>d26-d39</td><td>155.8</td><td>154.8</td><td>153.9</td><td>154.5</td><td>158.4</td><td>158.0</td><td>NS</td></tr><tr><td><strong>FC</strong></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>d0-d13</td><td>1.389<sup>a</sup></td><td>1.413<sup>ab</sup></td><td>1.480<sup>b</sup></td><td>1.371<sup>a</sup></td><td>1.394<sup>a</sup></td><td>1.388<sup>a</sup></td><td>0.009</td></tr><tr><td>d13-d26</td><td>1.462</td><td>1.473</td><td>1.510</td><td>1.503</td><td>1.487</td><td>1.432</td><td>NS</td></tr><tr><td>d26-d39</td><td>1.704</td><td>1.714</td><td>1.685</td><td>1.691</td><td>1.671</td><td>1.726</td><td>NS</td></tr><tr><td>WAFC<sub>1500</sub></td><td>1.524<sup>a</sup></td><td>1.539<sup>ab</sup></td><td>1.608<sup>b</sup></td><td>1.535<sup>ab</sup></td><td>1.522<sup>a</sup></td><td>1.493<sup>a</sup></td><td>0.002</td></tr></tbody></table>

Tr = treatment; BW = body weight; FI = feed intake; FC = feed conversion, WAFC<sub>1500</sub> = weight adjusted feed conversion = FC to obtain a body weight of 1500g, Tr 1 = normal calcium (Ca) and normal phosphorus (P) level, Tr 2 = low Ca and low P level, Tr 3 = normal Ca and low P level + normal phytase level, Tr 4 = normal Ca and low P level + high phytase level, Tr 5 = low Ca and low P level + normal phytase level, Tr 6 = low Ca and low P level + high phytase level

<sup>a-c</sup> means with the same letter are not significantly different from each other at P ≤ .05.

Daily feed intake was significantly lower for the PC (Tr1) group during the starter phase, however during the grower phase the NL dietary treatment group (Tr3) had the significantly lowest feed intake. Broilers fed the low Ca, low P (NC) diet had a feed intake comparable with the other dietary treatments. Adding both levels of phytase to the diets increased the feed intake of the broilers but not significantly with exception of the imbalanced diet during d21 to d26. Dietary treatments did not significantly influence feed intake during the finisher phase. For the entire period, broilers fed Tr3 and Tr5 had the lowest and highest feed intake, respectively.

During the starter period there was a significant effect of diets on feed conversion, however during grower and finisher this effect was no longer observed. During the starter period, the most favourable feed conversion was obtained if broilers were fed the imbalanced diet with a high dosage of phytase (Tr4) or the low Ca low P diet supplemented with phytase (Tr5 and Tr6). Due to a higher feed intake, feed conversion of Tr5 was comparable with Tr4. Feed conversion was significantly lower for the Tr3 fed broilers due to a low body weight gain. Supplementing a higher phytase level in combination with the imbalanced diet significantly improved feed conversion.
Broilers fed the negative control group without phytase supplementation (Tr2) had a significantly lower tibia ash percentage compared to the birds of the other dietary treatments and the highest prevalence of footpad lesions was seen at broilers fed the imbalanced diets.

## Conclusions

It can be concluded that a reduction of the minerals should be done in a balanced way to obtain maximal performance results. However, this study indicates that a higher dosage of phytase can compensate for these shortcomings. Furthermore, reducing the Ca and P level in a balanced way resulted in comparable results as the positive control group, however the significantly lower tibia ash % can be an indication of too low levels of both minerals in this study. Supplementing phytase did increase performance and the obtained results were even better than those of the PC group, but not at the 5% significance level. The higher dosage of phytase combined with the low Ca and P diet did not further improve the zootechnical results of the broilers.

## Literature

L'ETOURNEAU-MONTMINY, M. P., A. NARCY, P. LESCOAT, J. F. BERNIER, M. MAGNIN, C. POMAR, Y. NYS, D. SAUVANT, AND C. JONDREVILLE, 2010. Meta-analysis of phosphorus utilisation by chicks: Influence of dietary calcium and microbial phytase 555 content. Anim. 4:1844–1853
P-172 (ID 291)

EFFECTS OF DIETARY COARSENESS AND WET FEEDING ON P-UTILISATION OF LOW AVAILABLE P DIETS IN
YOUNG (PRE-)LAYER HENS

M.L. Staats-Elling<fnref n="1" />, K.A. Akinloye<fnref n="1" />, J. Loeffen<fnref n="1" />, Y. Yen<fnref n="1" />, A. Dijkslag<fnref n="2" />, R.P. Kwakkel<fnref n="1" />

**Abstract**

An experiment was conducted to study phosphorus utilization in 624 White Leghorn hens (15 to 28 wks) as affected by diets differing in particle size distribution ('feed structure'), conformation ('wet vs dry feed') and available phosphorus content in a 2x2x2 factorial design. Eight treatments were tested in 48 pens (6 replicate pens per treatment): a fine or a coarse mash diet, that was either fed in wet (1kg feed : 1L water) or dry form, and contained either a low (1.6g/kg) or standard (2.9g/kg) level of available phosphorus (aP). Experimental diets were maize based and contained no phytase supplements. The hypothesis was that a coarse and/or wet diet improves gastrointestinal tract development and alleviates the detrimental effects of a P-deficient diet by increasing the degradation of phytate. The hens were fed *ad libitum* provided in one meal per day. Feed intake (g/b/d) was increased when hens were fed the wet diet as compared to the dry diet in wk 19-22 (101.7 g vs. 98.7 g, P<0.001) and in wk 23 to 27 (116.0 g vs. 113.5 g, P<0.001). Feed structure affected intake in wk 19 to 22 (101.0 g fine diet vs. 99.4 g coarse diet; P=0.011) and in wk 23 to 27 (115.5 g fine diet vs. 114.1 g coarse diet; P=0.012). Rate of lay was highest for hens fed the combination of the wet and fine diet (93.01% vs. 90.17%; P=0.016). The coarse diet reduced egg mass production (49.69 g/d vs. 50.92 g/d, P=0.009). Tibia ash and mineral content and phytase activity in the GI-tract will be measured in due time and presented at the conference.

**Keywords:** young layer, diet structure, wet feed, phosphorus utilisation, performance

**Introduction**

A large majority of the phosphorus (P) found in plant ingredients of poultry feeds is bound to phytate, rendering this P
less available to the bird, unless broken down by the enzyme phytase. Despite improvements with the addition of mi-
crobial phytase, certain amounts of inorganic phosphates are still required in the diet. Hence the need to investigate
possible feeding strategies to improve P utilisation in poultry is evident. Providing birds with coarsely ground diets (lar-
ger particle sizes) and feeding it in wet form have shown to be beneficial to poultry. It has been demonstrated that feed-
ing broilers a coarse corn mash diet increased bone ash and P and Ca retention (Kasim and Edwards, 2000) compared
to those fed a fine corn mash diet, indicating an improved utilisation of P possibly due to an increased degradation of
phytate P. Wet feeding improves feed intake and digestibility (Yasar and Forbes, 1999). If and to what extend wet feed-
ing would improve the activity of phytase in the gut is unknown. It has, however, been shown that wetting feed reduces
phytate by fermentation, hence increasing the level of available P (aP) in the diet (e.g.: Blaabjerg et al. (2014)). This
study was aimed to investigate the effects of a coarse mash diet vs. a fine mash diet in combination with feeding the
diet soaked or dry on egg production, bone health and gastro-intestinal development in young layers fed a normal or
deficient level of calculated aP. It is hypothesized that coarse and wet feeding would counteract the negative effects of
a deficient aP level.

**Material and Methods**

A total of 624 110-day of White Novogen pullets were assigned to 8 treatment groups, in a 2x2x2 factorial design. Each treatment group consisted of 6 replicate pens, with 13 birds per pen. Dietary treatments consisted of a low P (1.6 g aP/kg diet) versus high P level (2.9 g aP/kg diet), dry versus wet feeding method and fine versus coarse mash diet. Diets were maize based, no phytase supplements were used and had a similar ingredient composition. Anorganic phosphate (MCP) was used to upgrade the low aP diets to the high aP diets. Diets were ground using a roller mill, with a roller distance of 2.3 mm for the fine diets and a distance of 3.4 mm for the coarse diets. The wet diets were prepared fresh daily prior to feeding by mixing the feed with tap water in a 1:1 ratio. The hens were fed *ad libitum* provided in one meal per day. Feed intake and egg production were recorded weekly. At 22 and 28 wks of age two hens per replicate were culled for GI-tract measurements and tibia collection. Chyme was collected from the crop, proventriculus and gizzard and small intestine, used for pH measurements and stored for later analysis of phytase activity. Empty organs

<footnote n="1">¹Animal Nutrition Group, Wageningen University, Wageningen, the Netherlands</footnote>

<footnote n="2">²Nutrition Innovation Centre, For Farmers, Lochem, the Netherlands</footnote>
were weighed. For the measurement of tibia breaking force a three point bending method was used, with the Instron 5564 texture analyser (compression speed was 2mm/min). Results are presented as mean ± SEM.

## Results

Feed intake (of the air-dry feed) was increased by wet feeding compared to dry feeding in the pre-layer period (wk 16-17; 87.3 g vs. 79.5 g, P<0.001) and in the transition period (wk 19-22; 101.7 g vs. 98.7 g, P<0.001). Feed intake in the transition period was also increased by the high P level compared to the low P level (101.0 g vs. 99.5 g, P=0.010) and the fine diet compared to the coarse diet (101.1 g vs. 99.4 g, P=0.011). FCR was unaffected by treatments. Rate of lay in the transition period was 68.43% and did not differ between treatments. Egg mass production was increased by the high P level compared to the low P level (33.07 g/d vs. 31.76 g/d, P=0.025) and by wet feeding compared to dry feeding (32.91 g/d vs. 31.73 g/d, P=0.035).

<table><caption>**Table 1:** Production performance measurements of hens fed diets differing in P level (P), conformation (C) and structure (S).</caption><thead><tr><th rowspan="3">P</th><th rowspan="3">C</th><th rowspan="3">S</th><th colspan="5">Production performance</th></tr><tr><th rowspan="2">Feed intake<br>g/d</th><th rowspan="2">FCR<sup>1</sup></th><th colspan="3">Age 23 - 27 wk</th></tr><tr><th>Rate of lay<br>%</th><th>Egg mass<br>g/hen/d</th><th>Egg weight<br>g/egg</th></tr></thead><tbody><tr><td rowspan="4">High P</td><td rowspan="2">Dry</td><td>Fine</td><td>112.7 ± 0.7</td><td>2.22 ± 0.02</td><td>91.08 ± 0.91</td><td>50.69 ± 0.44</td><td>55.97 ± 0.29</td></tr><tr><td>Coarse</td><td>113.4 ± 0.7</td><td>2.26 ± 0.03</td><td>91.99 ± 0.96</td><td>50.14 ± 0.53</td><td>54.97 ± 0.30</td></tr><tr><td rowspan="2">Wet</td><td>Fine</td><td>120.3 ± 0.6</td><td>2.29 ± 0.03</td><td>93.72 ± 0.89</td><td>52.58 ± 0.59</td><td>56.31 ± 0.26</td></tr><tr><td>Coarse</td><td>113.1 ± 0.7</td><td>2.33 ± 0.03</td><td>87.35 ± 1.20</td><td>48.36 ± 0.71</td><td>55.76 ± 0.31</td></tr><tr><td rowspan="4">Low P</td><td rowspan="2">Dry</td><td>Fine</td><td>112.9 ± 0.9</td><td>2.29 ± 0.04</td><td>89.61 ± 1.27</td><td>49.37 ± 0.86</td><td>55.40 ± 0.31</td></tr><tr><td>Coarse</td><td>115.1 ± 1.3</td><td>2.34 ± 0.03</td><td>89.45 ± 1.36</td><td>49.22 ± 0.74</td><td>55.19 ± 0.24</td></tr><tr><td rowspan="2">Wet</td><td>Fine</td><td>115.9 ± 0.7</td><td>2.27 ± 0.03</td><td>92.29 ± 1.13</td><td>51.05 ± 0.61</td><td>55.49 ± 0.25</td></tr><tr><td>Coarse</td><td>114.8 ± 0.7</td><td>2.26 ± 0.04</td><td>91.51 ± 1.36</td><td>51.02 ± 0.44</td><td>55.80 ± 0.21</td></tr><tr><th colspan="8" style="text-align: left;">Significance</th></tr><tr><td colspan="2">P level (P)</td><td></td><td>0.715</td><td>0.396</td><td>0.695</td><td>0.540</td><td><strong>0.039</strong></td></tr><tr><td colspan="2">Conformation (C)</td><td></td><td><strong>&lt;0.001</strong></td><td>0.547</td><td>0.402</td><td>0.066</td><td><strong>0.003</strong></td></tr><tr><td colspan="2">Structure (S)</td><td></td><td><strong>0.012</strong></td><td>0.095</td><td>0.051</td><td><strong>0.009</strong></td><td><strong>0.010</strong></td></tr><tr><td colspan="2">P * C</td><td></td><td><strong>0.035</strong></td><td><strong>0.005</strong></td><td><strong>0.040</strong></td><td>0.077</td><td>0.441</td></tr><tr><td colspan="2">P * S</td><td></td><td><strong>&lt;0.001</strong></td><td>0.612</td><td>0.167</td><td><strong>0.018</strong></td><td><strong>0.006</strong></td></tr><tr><td colspan="2">C * S</td><td></td><td><strong>&lt;0.001</strong></td><td>0.545</td><td><strong>0.016</strong></td><td>0.056</td><td>0.143</td></tr><tr><td colspan="2">P * C * S</td><td></td><td><strong>0.028</strong></td><td>0.436</td><td><strong>0.042</strong></td><td><strong>0.046</strong></td><td>0.900</td></tr></tbody></table>

Results from the early production period, wk 23-27, are shown in Table 1. There were no main treatment effects on rate of lay, but there were interaction effects. Hens fed the combination of the wet and fine diet had a higher rate of lay than hens fed the wet and coarse diet (93.01% vs. 89.43%, P=0.012), while no difference in rate of lay was found between the fine and coarse diets within the dry fed groups (90.35% and 90.72%). The hens fed the high P, wet and fine diet had a higher rate of lay than hens fed the high P, wet and coarse diet, but both groups did not differ from the other six groups. Egg mass production was reduced in hens fed the coarse diet compared to those fed the fine diet (49.69 g/d vs. 50.92 g/d, P=0.009). The hens fed the high P, wet and fine diet had a higher egg mass production than the hens fed the high P, wet and coarse diet and the hens fed the low P and dry diets, but did not differ from all other groups.

The average tibia breaking force was 132.33 N at wk 22 and 132.10 N at wk 28 and did not differ between groups. Relative empty gizzard weight was increased by the coarse diet compared to the fine diet at 22 wks of age (15.72 g/kg BW vs. 13.77 g/kg BW, P<0.001) and at 28 wks of age (14.32 g/kg BW vs. 13.51 g/kg BW, P=0.034). Empty gizzard weight was also increased by the low P diet compared to the high P diet at 22 wks of age (15.22 g/kg BW vs. 14.26 g/kg BW, P=0.010), but this effect was no longer significant at 28 wks of age. However, hens fed the low P and coarse diet had higher relative gizzard weights than hens fed the high P and coarse diet (15.08 g/kg BW vs. 13.56 g/kg BW, P=0.029), the low P and fine diet (13.34 g/kg BW, P=0.010) or the high P and fine diet (13.68 g/kg BW, P=0.049). Results for tibia ash and mineral content and phytase activity in the GI-tract are still pending.

<footnote n="1">¹ FCR= Feed conversion ratio (based on egg mass).</footnote>
Conclusions

Wet and coarse feeding did not counteract negative effects of the low P diet, as this diet had barely affected perform-
ance and had no effect on tibia strength, despite containing a level of only 1.6 g/kg aP. This could indicate that an aP
level of less than 2.9 g/kg is sufficient for maintaining egg production in young layers. Wet feeding increased feed intake
throughout the whole trial, similar to other findings (Yasar and Forbes, 1999). The coarse diet had negatively affected
feed intake, a finding which is in contrast to Safaa et al. (2009). Hens fed the combination of the wet and fine diet
showed the highest feed intake and egg performance. P level interaction effects for performance appear to be caused
by the contrast between the group fed the high P, wet and fine diet, which performed well, and the group fed the high P,
wet and coarse diet, which performed rather poorly. The coarse diet increased empty gizzard weight, similar to a num-
ber of other studies (Svihus, 2014). The positive effect of the low P diet on gizzard weight has, to the knowledge of the
authors, not been demonstrated before and could indicate an adaptive response.

Literature

BLAABJERG K, THOMASSEN AM AND POULSEN HD. 2014. Microbial phytase addition resulted in a greater increase in phosphorus digestibility in dry-fed compared with liquid-fed non-heat-treated wheat-barley-maize diets for pigs. Animal: 1-6.

KASIM AB AND EDWARDS HM. 2000. Effect of sources of maize and maize particle sizes on the utilization of phytate phosphorus in broiler chicks. Anim Feed Sci Technol **86**, 15-26.

SAFAA HM, JIMENEZ-MORENO E, VALENCIA DG, FRIKHA M, SERRANO MP AND MATEOS GG. 2009. Effect of main cereal of the diet and particle size of the cereal on productive performance and egg quality of brown egg-laying hens in early phase of production. Poult Sci **88**, 608-614.

SVIHUS B. 2014. Function of the digestive system. J Appl Poult Res **23**, 306-314.

YASAR S AND FORBES JM. 1999. Performance and gastro-intestinal response of broiler chickens fed on cereal grain-
based foods soaked in water. Br Poult Sci **40**, 65-76.
# THE EFFECT OF LIMESTONE PARTICLE SIZE AND PHYTASE SUPPLEMENTATION ON PERFORMANCE, EGG-SHELL QUALITY AND BACTERIAL SPECIES IN ILEUM

M. Englmaierová, M. Skřívan, and E. Skřívanová

Department of Nutrition Physiology and Animal Product Quality, Institute of Animal Science, v.v.i., Pratelstvi 815, 104 00 Prague-Uhrineves, Czech Republic

**Corresponding author:** englmaierova.michaela@vuzv.cz

## Abstract

The aim of the study was to examine the influence of limestone particle size and phytase (F) on performance of hens, physical parameters of eggs, digestibility of minerals and microbial colonization of the digestive tract. One hundred and sixty 60-week-old Lohmann Brown hens were housed in enriched cages for 12 months. The 2 x 2 factorial experiment included two levels of 3-phytase NATUPHOS® (0 and 350 FTU/kg) and two types of grain size of limestone (coarse limestone (CL) and mix of CL and fine limestone (FL) (65:35)) in mixed feed. All of the diets contained 4.4 g/kg of total phosphorus and 34 g/kg of calcium. Coarse limestone with F addition significantly increased hen-day egg production (P=0.018). The highest daily feed intake (P=0.001) and feed conversion (P=0.005) were recorded in the case of mix of CL and FL and F addition. The phytase supplement at 350 FTU/kg significantly increased shell percentage (P=0.007), shell index (P=0.003), shell strength (P=0.017) and shell thickness measured at blunt end (P=0.015), at the middle (P=0.009), at sharp end (P=0.015) and the average value of shell thickness (P=0.006). Albumen percentage and yolk percentage were influenced neither by limestone particle size nor by F supplementation. Albumen quality evaluated on the basis of Haugh units was higher (P=0.039) when using the mix of limestone. The phytase addition into the mixed feed nonsignificantly increased calcium and phosphorus digestibility in ileum by 6.2 and 9.1%, respectively. Bacteria belonging to genera *Turicibacter* and *Lactobacillus* occurred with higher frequency in ileum of hens fed with mixed feed enriched with the phytase.

**Keywords:** coarse limestone, 3-phytase, eggshell, microbial colonization, hen.

## Introduction

It is estimated that between 6 and 8% of the total eggs laid worldwide, are either cracked or broken before they reached the final consumer. Both in terms of economy and in relation to the health risks associated with a cracked shell, the eggshell quality still remains one of the main interests of the poultry industry. Coarsely ground limestone can increase the utilization of calcium (Ca) for shell formation and also for percentage a thickness of eggshell. Toward the end of the laying period, the utilization of Ca generally decreases. The utilization of Ca is 80% at the start of the laying cycle and 50% at the end of the laying cycle. It is assumed that coarsely ground limestone remains longer in the small intestine and Ca will be released during the night, when the shell is formed. Also phytase has positive effect on the shell quality. The addition of phytase completely overcomes the adverse effects associated with low intake of inorganic phosphorus (P) by animals and improves the bioavailability of Ca and eggshell quality at the marginal level of Ca 3.4%. In addition, phytase supplementation decrease P excretion in manure and reduce the potential environmental problems (Jalal and Scheideler, 2001). Exogenous phytases are mainly active in acidic pH which is in the crop, proventriculus and gizzard. There are several commercial phytase products such as 3-phytase NATUPHOS® produced by strain *Aspergillus niger*.

## Materials and Methods

One hundred and sixty 60-week-old Lohmann Brown hens were randomly assigned to 4 treatments with 4 replicate cages at 10 hens per cage. The hens were housed in three-floor enriched cages. The 2 x 2 factorial experiment was employed: 2 levels of phytase (F; 0 and 350 phytase units (FTU)/kg) and 2 types of grain size of limestone (coarse limestone (CL; 1 – 2 mm) and mix of CL and fine limestone (FL; < 0.5 mm) (65:35)) in mixed feed. NATUPHOS® (BASF, Ludwigshafen, Germany), a preparation of 3-phytase (EC 3.1.3.8) produced by *Aspergillus niger*, was chosen as the source of the F. The experiment lasted 12 months. All of the diets contained (per kg): approximately 11.5 MJ of apparent metabolizable energy (AMEN), 170 g of crude protein, 34 g of Ca and 4.4 g of total phosphorus. The number of eggs and hens and their health status were monitored daily. The hen-day egg production and feed intake were calculated weekly on a per-cage basis. Egg weights were determined once per week. For the physical parameter determination, eggs were collected in the 65th, 68th, and 70th week of the hens' age, and a total of 360 eggs were analyzed. The
shell breaking strength was determined on the vertical axis using an Instron 3360 apparatus (Instron, Canton, MA, USA). Haugh units and shell index were calculated according to the formulas. The shell thickness (values measured at the sharp and blunt ends and the equator, and the three obtained values were averaged) was measured using a micrometer after removing the shell membranes. Analyses of the P and Ca content of the eggshells were conducted once during the experiment in the 66th week of age of the hens (4 eggs per sample; n = 8). Dry samples of the eggshells were ashed at 550°C. The total P in the dried eggshells was assayed using a vanadate-molybdate reagent (AOAC, 2005; method 965.17). The calcium content was determined by atomic absorption spectrometry performed using a Solar M6 instrument (TJA Solutions, Cambridge, UK). Two groups of hens were used for the molecular analysis of microbial colonization (PCR-DGGE; n = 10) and digestibility determination (n = 10) in ileum: CL without and with F Natuphos® addition (350 FTU/kg). Digestibility determination using titanium dioxide as a marker was performed according to the modified methodology of Short et al. (1996). The resulting values were analyzed using Procedure of SAS (Statistical Analysis System, Version 9.2, 2003).

## Results

As is evident from Table 1, CL with F addition at 350 FTU/kg significantly (P=0.018) increased hen-day egg production (88.6%) contrary to alone CL (82.5%) or mix of CL and FL with F supplement (82.7%). Significantly lower daily feed intake (P=0.001) and feed conversion (P=0.005) were recorded in the hens fed with the mix of CL and FL without F addition and in the diet with CL together with F. The egg weight was not influenced.

<table><caption>**Table 1.** The effect of phytase and limestone particle size on performance</caption><thead><tr><th rowspan="2">Phytase (F; FTU/kg)<br>Limestone (L)</th><th colspan="2">0</th><th colspan="2">350</th><th rowspan="2">SEM</th><th colspan="3">Probability</th></tr><tr><th>FL+CL</th><th>CL</th><th>FL+CL</th><th>CL</th><th>F</th><th>L</th><th>F x L</th></tr></thead><tbody><tr><td>Hen-day egg production (%)</td><td>84.9<sup>ab</sup></td><td>82.5<sup>b</sup></td><td>82.7<sup>b</sup></td><td>88.6<sup>a</sup></td><td>0.88</td><td>NS</td><td>NS</td><td>0.018</td></tr><tr><td>Egg weight (g)</td><td>66.7</td><td>66.5</td><td>66.4</td><td>65.9</td><td>0.19</td><td>NS</td><td>NS</td><td>NS</td></tr><tr><td>Feed intake (g/day/hen)</td><td>116.2<sup>b</sup></td><td>116.9<sup>b</sup></td><td>122.4<sup>a</sup></td><td>116.3<sup>b</sup></td><td>0.57</td><td>0.009</td><td>0.012</td><td>0.001</td></tr><tr><td>Feed conversion ratio (g/g)</td><td>2.08<sup>b</sup></td><td>2.15<sup>ab</sup></td><td>2.30<sup>a</sup></td><td>2.03<sup>b</sup></td><td>0.030</td><td>NS</td><td>NS</td><td>0.005</td></tr></tbody></table>

FTU = phytase units; FL = fine limestone; CL = coarse limestone; NS = non-significant

<sup>a-b</sup> means in the same row with different superscripts differ significantly

As regards the physical parameters of egg quality (Table 2), F addition increased eggshell quality. The phytase supplement at 350 FTU/kg significantly increased shell percentage (P=0.007), shell index (P=0.003), shell strength (P=0.017) and shell thickness measured at blunt end (P=0.015), at the middle (P=0.009), at sharp end (P=0.015) and the average value of shell thickness (P=0.006). Limestone particle size influenced only Haugh units.

<table><caption>**Table 2.** The effect of phytase and limestone particle size on egg quality</caption><thead><tr><th rowspan="2">Phytase (F; FTU/kg)<br>Limestone (L)</th><th colspan="2">0</th><th colspan="2">350</th><th rowspan="2">SEM</th><th colspan="3">Probability</th></tr><tr><th>FL+CL</th><th>CL</th><th>FL+CL</th><th>CL</th><th>F</th><th>L</th><th>F x L</th></tr></thead><tbody><tr><td>Haugh units</td><td>79.8</td><td>79.9</td><td>82.7</td><td>77.4</td><td>0.69</td><td>NS</td><td>0.039</td><td>NS</td></tr><tr><td>Albumen percentage (%)</td><td>64.4</td><td>64.5</td><td>63.9</td><td>64.4</td><td>0.18</td><td>NS</td><td>NS</td><td>NS</td></tr><tr><td>Yolk percentage (%)</td><td>25.8</td><td>25.4</td><td>25.9</td><td>25.2</td><td>0.16</td><td>NS</td><td>NS</td><td>NS</td></tr><tr><td>Shell thickness</td><td></td><td></td><td></td><td></td><td></td><td></td><td></td><td></td></tr><tr><td>Blunt end (µm)</td><td>347</td><td>359</td><td>364</td><td>370</td><td>3.0</td><td>0.015</td><td>NS</td><td>NS</td></tr><tr><td>Equator (µm)</td><td>356</td><td>363</td><td>371</td><td>380</td><td>3.0</td><td>0.009</td><td>NS</td><td>NS</td></tr><tr><td>Sharp end (µm)</td><td>359</td><td>364</td><td>374</td><td>380</td><td>3.2</td><td>0.015</td><td>NS</td><td>NS</td></tr><tr><td>Average (µm)</td><td>354</td><td>362</td><td>370</td><td>377</td><td>2.8</td><td>0.006</td><td>NS</td><td>NS</td></tr><tr><td>Shell breaking strength (g/cm²)</td><td>3,893</td><td>3,826</td><td>4,165</td><td>4,320</td><td>78.4</td><td>0.017</td><td>NS</td><td>NS</td></tr><tr><td>Shell index (g/100 cm²)</td><td>8.6</td><td>8.7</td><td>8.9</td><td>9.1</td><td>0.06</td><td>0.003</td><td>NS</td><td>NS</td></tr><tr><td>Shell percentage (%)</td><td>9.8</td><td>10.0</td><td>10.2</td><td>10.4</td><td>0.07</td><td>0.007</td><td>NS</td><td>NS</td></tr></tbody></table>

FTU = phytase units; FL = fine limestone; CL = coarse limestone; NS = non-significant
Nonsignificant increase of Ca and P digestibility in ileum was ascertained after F addition into the mixed feed (Table 3). In addition, F supplement had an impact on microbial colonization of the digestive tract of hens. Bacteria belonging to genera *Turicibacter* and *Lactobacillus* occurred in ileum with higher frequency.

<table><caption>**Table 3.** The effect of phytase and limestone particle size on calcium and phosphorus digestibility in ileum</caption><thead><tr><th>Phytase (FTU/kg)</th><th>0</th><th>350</th><th>SEM</th><th>Probability</th></tr><tr><th>Limestone</th><th>CL</th><th>CL</th><th></th><th></th></tr></thead><tbody><tr><th>Calcium (%)</th><td>49.9</td><td>53.0</td><td>4.44</td><td>NS</td></tr><tr><th>Phosphorus (%)</th><td>42.9</td><td>46.8</td><td>2.32</td><td>NS</td></tr></tbody></table>

FTU = phytase units; CL = coarse limestone; NS = non-significant

## Conclusions

Feeding the hens a diet with coarse limestone with grain size of 1 – 2 mm, as a source of calcium, enriched with the 3-phytase at 350 FTU/kg leads to increase of performance. The addition of phytase alone improves eggshell quality, increases calcium and phosphorus digestibility and changes bacterial colonization of ileum.

## Acknowledgement

This study was supported by the Ministry of Agriculture of the Czech Republic (Projects No. QJ1310002 and No. MZE RO0714).

## Literature

AOAC, 2005: Official Methods of Analysis, 18th ed., AOAC International, Gaithersburg, MD, USA.

JALAL, M.A., S.E. SCHEIDELER, 2001: Effect of supplementation of two different sources of phytase on egg production parameters in laying hens and nutrient digestibility. Poult. Sci. **80**, 1463–1471.

SAS, 2003: SAS/STAT User's Guide (release 9.2). Cary, NC, USA, SAS Institute.

SHORT, F.J., P. GORTON, J. WISEMAN, K.N. BOORMAN, 1996: Determination of titanium dioxide added as an inert marker in chicken digestibility studies. Anim. Feed Sci. Technol. **59**, 215–221.
P-175 (ID 180)

# THE EFFECT OF CALCIUM AND HOUSING SYSTEM ON PERFORMANCE AND EGG COMPOSITION OF TWO CONTRAST GENOTYPES

M. Ketta, E. Tůmová

Dept. of Animal Husbandry, Czech University of Life Science, Prague, Czech Republic

Corresponding author: kettam@af.cz.u.cz

## Abstract

The objective of this study was to compare performance and egg composition of Lohmann White and Czech Hen housed in cages and on litter system in accordance to different calcium levels (3.0 vs. 3.5 %). Higher hen-day egg production (P≤0.001) and feed intake (P≤0.001) were observed in Lohmann White. Feed intake was significantly (P≤0.001) higher on litter (159.49 g/day) in comparison with cages (89.30 g/day). Egg weight was significantly (P≤0.001) affected by the interactions of all evaluated factors. The heaviest eggs (61.8 g) were detected in Lohmann White housed on litter with 3.5 % Ca and the lightest (47.3 g) were in Czech Hen housed in cage system with 3.0 % Ca. Significantly (P≤0.001) higher albumen percentage was observed in Lohmann White (61.1 %) than Czech Hen (57.6 %). Czech Hen produced eggs with significantly (P≤0.001) higher yolk percentage (26.3 %) than Lohmann White (22.7 %). Significantly (P≤0.001) higher eggshell percentage (12.2 %) was obtained in eggs from Czech Hen in comparison with Lohmann White (11.4 %). The performance and egg composition were affected by genotype and housing system; however, egg weight by interactions of all evaluated factors. Growing Ca content increased egg weight.

**Keywords:** Laying hen; genotype; housing; Ca; performance

## Introduction

The hen performance and egg quality parameters are affected by many factors including calcium content of feed, housing system and hen genotype. Ca requirements by laying hens are considered an ongoing challenge for nutritionists and poultry producers. SAFAA et al. (2008) indicated that, increasing Ca levels significantly improved egg production, egg weight and eggshell weight. The effect of housing system on hens performance were described by ENGL-MAIEROVÁ et al. (2014) who found lower feed consumption and higher egg production in conventional cages compared to litter system. Egg weight also varies according to different housing systems. SINGH et al. (2009) detected heavier eggs on litter, whereas, TŮMOVÁ et al. (2011) in cages. Depending on laying hen genotype variances, there are differences in hen performance. Higher egg production was found in commercial genotype compared to the non-commercial ones by SINGH et al. (2009). Egg quality characteristics including egg weight, yolk weight, albumen weight and albumen/yolk ratio differed between variable brown genotypes (TŮMOVÁ et al., 2011).

This study aimed on evaluating of hen performance and egg quality characteristics in Lohmann White and Czech Hen housed in cages and on litter with different feed calcium content.

## Material and Methods

Laying hens performance and egg quality parameters were evaluated in an experiment with 132 laying hens of Lohmann White and pure breed Czech Hen. The hens were housed in conventional cages Eurovent (72 hens, 550 cm² / hen) and on six littered pens (60 hens, 7 hens/m², 10 hens/pen). In both housing systems laying hens were fed feed mixtures differed in Ca content, K (control group) 3.48 % and P (experimental group) 3.0 %. The feed mixture between 20 to 40 weeks of age consisted of 16.66% crude protein (CP), 11.4 MJ of ME; and 15.37% CP, 11.48 MJ ME from 40 to 60 weeks of age.

Eggs for physical quality determination were collected in four week interval two days in row. Eggs were weighted after collection. Albumen and yolk weights were individually measured. Eggshell weight was determined after drying. The albumen, yolk, shell percentage and yolk/albumen ratio were calculated. Hen performance and egg quality data were evaluated by three-way analysis of variance using the GLM procedure (housing, genotype, calcium interactions) of SAS (SAS Institute Inc., Cary, Nc, 2003).
## Results

The hen performance parameters (Table 1) were significantly (P≤0.001) affected by genotype. Higher hen-day egg production and daily feed intake were revealed to Lohmann White in comparison with Czech Hen. Similar results were reported by SINGH et al. (2009) who found higher egg production from commercial genotype in comparison with the non-commercial ones. Significant effect of Ca level (P≤0.05) was observed in egg weight with heavier eggs at 3.5% than 3.0% Ca content, which corresponds with the results of NARVAEZ-SOLARTE et al. (2006) who found an increasing linear effect of Ca level on egg weight. The effect of housing system was detected in daily feed intake with higher daily consumption on litter than cages. Similarly, ENGLMAIEROVÁ et al. (2014) detected higher feed consumption on litter compared to conventional cages.

<table><caption>**Table 1.** Main effects of genotype, housing system, and Ca content on hen performance.</caption><thead><tr><th>Factor</th><th>Treatment</th><th>Ca</th><th>Hen-day egg production (%)</th><th>Feed intake (g)</th><th>Egg weight (g)</th></tr></thead><tbody><tr><td rowspan="6">Czech Hen</td><td rowspan="2">Cage</td><td>3.00</td><td>19.18</td><td>65.12</td><td>47.34<sup>e</sup></td></tr><tr><td>3.50</td><td>13.23</td><td>65.38</td><td>51.63<sup>cd</sup></td></tr><tr><td rowspan="2">Litter</td><td>3.00</td><td>21.30</td><td>150.70</td><td>50.91<sup>d</sup></td></tr><tr><td>3.50</td><td>19.15</td><td>143.70</td><td>48.20<sup>cd</sup></td></tr><tr><td rowspan="6">Lohmann White</td><td rowspan="2">Cage</td><td>3.00</td><td>88.20</td><td>116.06</td><td>60.86<sup>ab</sup></td></tr><tr><td>3.50</td><td>81.03</td><td>110.62</td><td>60.45<sup>b</sup></td></tr><tr><td rowspan="2">Litter</td><td>3.00</td><td>86.90</td><td>169.00</td><td>58.44<sup>c</sup></td></tr><tr><td>3.50</td><td>89.55</td><td>174.55</td><td>61.83<sup>a</sup></td></tr><tr><td>RMSE</td><td></td><td>11.17</td><td>10.31</td><td>6.04</td></tr><tr><td>Genotype</td><td></td><td>0.001</td><td>0.001</td><td>0.001</td></tr><tr><td>Housing</td><td></td><td>0.927</td><td>0.001</td><td>0.001</td></tr><tr><td>Calcium</td><td></td><td>0.959</td><td>0.560</td><td>0.033</td></tr><tr><td>Genotype x Housing x Ca</td><td></td><td>0.784</td><td>0.365</td><td>0.001</td></tr></tbody></table>

Significant interaction between genotype, housing system and Ca levels was detected in egg weight. The heaviest eggs (61.83 g) were found in Lohmann White housed on litter with 3.5% Ca, while, the lightest eggs (47.34 g) were in Czech Hen housed in cages with 3.0 % Ca. Egg quality parameters were affected mainly by genotype (Table 2). Significantly (P≤0.001) higher albumen percentage and shell percentage was found in eggs from Lohmann White in comparison with Czech Hen. On the other hand, eggs from Czech Hen showed higher values of yolk percentage and albumen/yolk ratio than Lohmann White genotype. These results in agreement with the founding of (TŮMOVÁ et al., 2011) who reported significant differences in yolk weight, albumen weight and albumen/yolk ratio between variable brown genotypes. Increasing Ca levels significantly (P≤0.009) increased eggshell percentage which correspond

<footnote>a, b, c, d,e statistically significant differences (P≤0.01) within columns are indicated by different superscripts.</footnote>
<table><caption>**Table 2.** Main effects of genotype, housing system, and Ca content on egg quality parameters.</caption><thead><tr><th>Factor</th><th>Treatment</th><th>ca</th><th>Albumen percentage (%)</th><th>Yolk percentage (%)</th><th>Shell percentage (%)</th><th>Albumen / yolk ratio</th></tr></thead><tbody><tr><td rowspan="4">Czech Hen</td><td rowspan="2">Cage</td><td>3.0</td><td>57.59</td><td>28.46</td><td>11.62</td><td>53.77</td></tr><tr><td>3.5</td><td>57.21</td><td>26.58</td><td>10.87</td><td>56.25</td></tr><tr><td rowspan="2">Litter</td><td>3.0</td><td>57.16</td><td>24.15</td><td>11.84</td><td>54.64</td></tr><tr><td>3.5</td><td>58.40</td><td>26.11</td><td>11.39</td><td>51.99</td></tr><tr><td rowspan="4">Lohmann White</td><td rowspan="2">Cage</td><td>3.0</td><td>61.46</td><td>22.94</td><td>12.17</td><td>43.19</td></tr><tr><td>3.5</td><td>61.32</td><td>22.72</td><td>12.45</td><td>42.97</td></tr><tr><td rowspan="2">Litter</td><td>3.0</td><td>60.44</td><td>22.34</td><td>12.05</td><td>45.81</td></tr><tr><td>3.5</td><td>61.30</td><td>22.68</td><td>12.28</td><td>43.35</td></tr><tr><td>RMSE</td><td></td><td></td><td>2.62</td><td>6.74</td><td>0.890</td><td>6.41</td></tr><tr><td>Genotye</td><td></td><td></td><td>0.001</td><td>0.001</td><td>0.001</td><td>0.001</td></tr><tr><td>Housing</td><td></td><td></td><td>0.072</td><td>0.092</td><td>0.314</td><td>0.055</td></tr><tr><td>Calcium</td><td></td><td></td><td>0.015</td><td>0.878</td><td>0.009</td><td>0.002</td></tr><tr><td>Genotye x Housing x Ca</td><td></td><td></td><td>0.425</td><td>0.099</td><td>0.189</td><td>0.126</td></tr></tbody></table>

with SAFAA et al. (2008) who reported an increasing in eggshell percentage with increasing Ca levels.

## Conclusions

Results of the study show that only egg weight was affected by interaction of all evaluated factors. Better performance, higher egg weight and albumen proportion in Lohmann hen is the result of the long term selection. There was a minor effect of housing system on proportion of egg components but Ca had a higher impact on egg composition.

## Acknowledgement

The study was supported by the Ministry of Agriculture of the Czech Republic, a project of NAAR No. QJ1310002.

## Literature

ENGLMAIEROVÁ, M., E. TŮMOVÁ, V. CHARVATOVÁ, M. SKŘIVAN, 2014: Effects of laying hens housing system on laying performance, egg quality characteristics, and egg microbial contamination. Czech. J. Anim. Sci. **59**, 345-352.

NARVAEZ-SOLARTE, W., H.S. ROSTAGNO, P.R. SOARES, L.F. URIBE-VELASQUEZ, M.A. SILVA M.A, 2006: Nutritional requirement of calcium in white laying hens from 46 to 62 week of age. Int. J. Poult. Sci. **5**, 181-184.

SAFAA, H.M., M.P. SERRANO, D.G. VALENCIA, M. FRIKHA, E. JIMÉNEZ-MORENO, G. G. MATEOS, 2008: Productive performance and egg quality of brown egg-laying hens in the late phase of production as influenced by level and source of calcium in the diet. Poult. Sci. **87**, 2043-2051.

SINGH, R., K.M. CHENG, F.G. SILVERSIDES, 2009: Production performance and egg quality of four strains of laying hens kept in conventional cages and floor pens. Poult. Sci. **88**, 256-264.

TŮMOVÁ, E., M. ENGLMAIEROVÁ, Z. LEDVINKA, V. CHARVATOVÁ, 2011: Interaction between housing system and genotype in relation to internal and external egg quality parameters. Czech. J. Anim. Sci. **56**, 490-498.
# EFFECT OF FORM OF SELENIUM USED BROILER BREEDERS' DIET ON CHICKS' GROWTH PERFORMANCE

H.R.Kutlu, L.Çelik, N. Saber, H.Cinli, F. Küley, M. Kutlu, O. Yücelt

University of Çukurova, Turkey

## Abstract

The present study was conducted to evaluate performance of broilers chicks obtained from breeders receiving two different sources (Se-hydroxy-4-methylselenobutanoic acid; HMSeBA vs. selenium yeast) of organic selenium. Two hundred and twenty, 54 weeks old Ross-308 male (20) and female (200) breeders were fed standard breeder (female) diets containing organic selenium sources; HMSeBA or Selenium yeast from 54 to 62 weeks of age. For hatchings process, eggs were collected, selected and stored in a cool room until owing 700 eggs, 350 from each group (35 x 10 subgroup) between 58-59 weeks of age for the first hatched and 61-62 weeks of age for the second hatched, respectively. Eggs were then incubated in a single stage incubator using 35 eggs per replication in order to obtained chicks to assess effect of selenium sources in maternal diets on growth performance of progeny. Following hatchings, chicks were weighed than accommodated in 20 pens, each treatment group had 10 pens, each sized 2x3m and equipped with a tube feeder and water drinkers on litter; wood shaving litter 7-8 cm height. Chicks were fed ad libitum with standard starter, grower, finisher and withdrawal diets containing no organic selenium source but inorganic selenium for a period of 35 days under a 23:1 light: dark photoperiod. Chick's performance was assesed by measuring weigh, weigh gain, feed intake and feed conversion efficiency. At 35 days of age birds were weighed and transferred to slaughterhouse. The data obtained in the experiment were analysed using t-test.

The results obtained during the first performance tests showed that chicks from maternal receiving HMSeBA had higher body weight gain at 7 and 14 days of age (P<0.05). At 35 days of age, the chicks obtained from the breeder receiving HMSeBA had numerically higher (P>0.05) body weight, feed intake and better feed efficiency (P>0.05) than those of maternal receiving selenium yeast. The results of the second performance test revealed only numerically difference (P>0.05) in the favour of chicks obtained from the breeders receiving HMSeBA. However, the performance values obtained from the both groups were found to be greatly higher than breeder's expectation in the both trials. All these results may suggest that providing HMSeBA to breeder diets could be of value increasing progeny performance in terms of growth performance with a higher efficacy.

**Keywords:** Organic Selenium, Breeder, Egg, Hatching, Broiler, Performance

## Introduction

It is well documented that selenium with the conjunction of vitamin E is the vital nutrient source to maintain health, growth and product quality in mammals and birds (1). Vitamin E is a fat-soluble nutrient found in body fat depots, plasma lipoproteins, and cell membrane phospholipids, where it serves as an important antioxidant (2). Selenium (Se) fulfils an antioxidant role as a component of glutathione peroxidises (GSHPx). Se is widely distributed in the body, but the most labile reservoir is in the liver (2). It is found in body tissue principally as selenomethionine (SeMet) or as selenocysteine (SeCys), the latter found in GSHPx. Both nutrients are should be examine together because of their related functions and similar deficiency signs. However, Se and vitamin E each have unique metabolic roles, and the factors that alter the oxidative state of an animal may differentially affect their dietary needs. It is now well known that Se plays an important role in maintaining semen quality (3). An optimal Se status in male birds is considered to be an important factor in ensuring the fertility of breeding stocks, while an optimal Se status of the eggs of females needs for antioxidant system to maintain embryo development and also hatchability (4).

In poultry nutrition, selenium requirements meet through trace mineral premixes. Recent literature showed that besides the rational source of supplemental selenium in mineral forms mainly sodium selenite in animal diets, organic selenium sources have also been developed through selenize yeast with Se in the form of selenomethionine and also 2-hydroxy-4-methylselenobutanoic acid (HMSeBA) in recent years (5). Comparative studies showed that organic Se source provides some advantages with higher bioavailability and lower inclusion rate in the diet (5). However no comparative studies have been undertaken to evaluate different sources of organic selenium in female broiler breeders in terms of reproduction performance, egg quality, hatchability and chicks' performance.
The present project was aimed to evaluate organic sources of Se-hydoxy-4-methylselenobutanoic acid (HMSeBA of Adisseo; Selisseo) in contrast to another organic selenium source (selenium yeast) used in broiler breeders' diet on chicks' growth performance.

## Materials and Methods

The present study was carried out in the Broiler Breeder Unit of the Experimental Farm of the Department of Animal Science, Faculty of Agriculture, University of Çukurova, Adana-Turkey. Two hundred and twenty, 54 weeks of age Ross-308 male (20) and female (200) breeders were used in the experiment lasted 9 weeks. Throughout the experimental study standard breeder (female) diet based on corn and soya was used supplemented with selenium in the form of SE-yeast or HMSeBA at 200 ppb.

During the experiment, hatching performance of the eggs obtained from the birds receiving HMSeBA or Se-yeast was performed by 21 day-incubation twice. For hatchings process, egg selected and stored in a cool room until owing 700 eggs, 350 from each group (35x10 subgroups) between 58-59 weeks of age for the first hatched and 61-62 weeks of age for the second hatched, respectively. Eggs were incubated in a single stage incubator using 35 eggs per replication. Temperature was controlled according to McQuoid (6). Hatching was completed on the 21st day. The chicks were carefully removed from the pouches and their weights were determined by using an electronic scale with a sensitivity of 0.01 g. Chicks were then accommodated in 20 pens, each treatment groups has 10 pens sized 2x3m and equipped with a tube feeder and water drinkers on litter; wood shaving litter 7-8 cm height. Chicks were fed 35 days ad libitum under a 23:1 light: dark photoperiod. The chicks obtained from the eggs of a group/subgroup were housed by name of the maternal group/subgroup number to fellow on the same axis to maternal to its chicks. Chicks were received four stages (starter; CP:24%, ME:3050 Kcal/kg, grower; CP:22%, ME:3150 Kcal/kg, finisher; CP:21%, ME:3250 Kcal/kg, and withdrawal; CP:20%, ME:3250 Kcal/kg, feeds) feeding programme with the diets containing conventional vitamins and trace mineral premixes, with 150 ppb selenium as sodium selenite. During the experiment, live weight, feed intake and feed efficiency were recorded weekly in subgroup (10 replicates in each group) level. At 35 days of age, birds were attained the weight given for 42 days of age by the Breeder Handbook (7), therefore feeding was ceased and birds were taken slaughterhouse one week earlier. The data obtained in the study were analysed using t-test procedure of SAS; the Statistical Analysis System (8). Result obtained in this study will be presented as means per bird with standard errors of the difference between means (SED) and CV.

## Results and Discussion

At the end of the first and the second incubations, all the chicks obtained at hatching were subjected to growth performance test by feeding starter, grower, finisher and withdrawal feeds containing no organic selenium source but inorganic selenium (sodium selenite) for a period of 35 days during which all the animals were allocated in a group named by its maternal group. When the project was proposed in the first place, broiler performance tests at the end of the both incubation were planned to carry out 42 days, however, in order to control leg problems due to enormously fast growing during the testing period did not let trials to be run 42 days. At 35 days of age in the first and the second trials broilers had to be taken slaughterhouse. The results are given here were obtained at 35 days of age in the first and the second performance test.

The results obtained during the first performance test were given in Table 1. The chicks of the both groups grew very fast. Especially chicks from maternal receiving HMSeBA had mean slaughter weight of 2670 grams at 35 days of age, being a 7-days earlier and 0.09 better feed conversion rate (1.52) than breeder's expectation (1.61) at 35 days of age. Chicks from maternal receiving Se-Yeast had mean slaughter weight of 2606 grams at 35 days of age, being a 5-days earlier and 0.10 better feed conversion rate (1.51) than breeder's expectation (1.61) at 35 days of age. The results also showed that at the end of the feeding period, the chicks, whose maternal received HMSeBA attained body mass almost 6% higher than those of the birds of Se-Yeast group. The both groups had similar liveability, feed intake and also feed conversion rate (Tables 1 and 2) throughout the feeding period.

The results obtained during the second performance test were given in Table 3. The chicks of the both groups grew very fast similar to the first performance test. Especially chicks from maternal receiving HMSeBA had mean slaughter weight of 2531 grams at 35 days of age, being a 6-days earlier and 0.11 better feed conversion rate (1.49) than breeder's expectation (1.61) at 35 days of age. Chicks from maternal receiving Se-Yeast had mean slaughter weight of 2481 grams at 35 days of age, being a 4-days earlier and 0.06 better feed conversion rate (1.55) than breeder's expectation (1.61) at 35 days of age. The results also showed that at the end of the feeding period, the