Fatty acid composition, physicochemical and sensory properties of

Europ.Poult.Sci., 78. 2014, ISSN 1612-9199, © Verlag Eugen Ulmer, Stuttgart. DOI: 10.1399/eps.2014.61


Fatty acid composition, physicochemical and sensory properties of eggs from laying hens fed diets containing blue lupine seeds Fettsäuremuster, physikochemische und sensorische Eigenschaften von Eiern nach Fütterung der Legehennen mit Rationen mit blauen Lupinen 1

1

2

2

3

Aleksandra Drażbo , D. Mikulski , Z. Zdunczyk , Beata Szmatowicz , A. Rutkowski and J. Jankowski

1

1

Department of Poultry Science, University of Warmia and Mazury, Olsztyn, Poland

2

Institute of Animal Reproduction and Food Research, Polish Academy of Sciences, Olsztyn, Poland

3

Department of Animal Nutrition and Feed Management, PoznanUniversity of Life Sciences, Poznań, Poland

Correspondence: [email protected] Manuscript received 23 March 2014, accepted 14 June 2014

Introduction Soybean meal, the most common vegetable protein source in poultry diets is rich in protein with the desired amino acid composition (BAKER, 2000). Increased worldwide demand for feed protein from soybean drives up its prices (PETTERSON, 2000; LAUDADIO and TUFARELLI, 2010). For both economic and social reasons, efforts are being made to totally or partially replace soybeans and soybean meal with local (European) protein sources in livestock diets (SUCHÝ et al., 2006, JANKOWSKI et al., 2011; MIKULSKI et al., 2012) In layer diets, lupine seeds can only be used as a partial substitute for soybean meal, due to their lower protein and metabolizable energy content, and the presence of anti-nutritional compounds (RUBIO et al., 2003), which is the major factor limiting the use of legumes in poultry diets (OLKOWSKI et al., 2005). Breeding low-alkaloid lupine varieties and inactivation or removal of anti-nutritional factors from lupine seeds contributed to the use of legumes as an alternative for conventional protein sources in poultry nutrition (LAUDADIO and TUFARELLI, 2011). Apart from low levels of anti-nutritional factors, modern lupine varieties are also characterized by higher concentrations of high-quality protein, a higher yield and a shorter growing season (PRUSIŃSKI, 2007). Blue lupine,

compared with other lupine species, has a higher yield potential, can tolerate a wider range of temperatures, and grows faster (WIATR et al., 2007).

The seeds of blue lupine can be a valuable ingredient of laying hen diets due to their low alkaloid levels (MARTÍNEZVILLALUENGA et al., 2006) and high concentrations of unsaturated fatty acids (UFAs) (BOSCHIN et al., 2008; RESTA et al., 2008). The inclusion of lupine in layer diets may increase the nutritional value of eggs (STRAKOVÁ et al., 2010). Dietary modifications of the fatty acid profile of yolk lipids and poultry meat have been studied extensively (ZDUŃCZYK and JANKOWSKI, 2013). Blue lupine seeds, used as a substitute for soybean meal, could also enrich the egg yolk with essential UFAs. The use of blue lupine seeds as a protein source in layer diets remains insufficiently researched. Therefore, the objective of this study was to determine the effect of different blue lupine content of layer diets on the physicochemical and sensory properties of eggs, and the fatty acid profile of yolk.

25.08.2014

1 / 14


Materials and Methods

Birds The experimental materials comprised 150 Lohmann Brown laying hens aged 18 weeks. Before the experiment, all

birds were weighed individually and were placed in individual cages (30 × 60 cm) where they were kept during the first 20 weeks of the laying period (from 18 to 38 weeks of age) under controlled environmental conditions, with 14 hours light and 10 hours dark. The replicates were equally distributed among three cage levels to minimize the cage

level effect. All birds had free access to feed and water. The protocol for this study was approved by the local Animal Experimentation Ethics Committee.

Diets Seeds of blue lupine (Lupinus angustifolius cv. Sonet), purchased from the Plant Breeding Station in Poznań, were used in the study. This cultivar is characterized by a low alkaloid content of seeds and a high dry matter protein

content. Laying hens were randomly assigned to three dietary treatments (50 replicates in each, 1 hen = 1 replicate), and were fed diets containing 16.5% crude protein and 11.3 MJ/kg AME as per nutrient requirements of hens (SMULIKOWSKA and RUTKOWSKI, 2005). Iso-protein and iso-energetic diets differed in protein sources, as follows:

group C – soybean meal and rapeseed meal; group L10 – soybean meal, rapeseed meal and 10% blue lupine; group L20

– soybean meal, rapeseed meal and 20% blue lupine. All diets contained identical concentrations of lysine, methionine plus cystine, and minerals (Ca and P). The apparent metabolizable energy (AME) content of blue lupine seeds was assumed at 7.22 MJ/kg based on the Polish feedstuff analysis tables (SMULIKOWSKA and RUTKOWSKI, 2005). The composition and nutritional value of experimental diets, and the chemical composition of blue lupine seeds are shown in Tables 1 and 2.

25.08.2014

2 / 14


Table 1. Composition and nutritional value of diets fed to laying hens (% as-fed basis) Zusammensetzung und Nährstoffgehalte der Versuchsrationen (% Frischsubstanz)

Diet1

Composition, %

C

L10

L20

Wheat Triticale Soybean meal Rapeseed meal

59.14 10.00 14.51 4.00

51.32 10.00 10.40 4.00

43.51 10.00 6.28 4.00

Blue lupine Soybean oil and lard (1:1) Salt Limestone Monocalcium phosphate

– 2.04 0.37 8.85 0.37

10.00 3.92 0.38 8.81 0.42

20.00 5.82 0.38 8.77 0.46

0.13

0.14

0.16

0.09

0.11

0.12

0.50

0.50

0.50

AME. MJ/kg

11.3

11.3

11.3

Total protein Crude fat Acid Detergent Fiber (ADF)

165 36.0 39.2

165 57.1 54.5

165 78.4 70.5

Neutral Detergent Fiber (NDF) Raffinose family oligosaccharides (RFOs) Lysine Methionine Met + Cys Arginine

104.9 7.7 7.8 3.8 7.0 9.4

115.2 14.2 7.8 3.6 7.0 10.4

125.4 20.8 7.8 3.5 7.0 11.3

Threonine Tryptophan

5.5 2.0

5.5 1.9

5.5 1.8

Ca Non-phytin P

36.0 2.5

36.0 2.5

36.0 2.5

DL-Methionine2 3

L-Threonine Vitamin-mineral premix Nutritional value4, g/kg

1

C – control diet, L10 – 10% of blue lupine in the diet, L20 – 20% of blue lupine in the diet MetAMINO®, Evonik Degussa Gmbh, Essen, Germany, 990 g/kg of methionine. 3 Ajinomoto Eurolysine S.A.S, Amiens, France, 780 g/kg of lysine 4 Calculated from the analyses of feed ingredients – data provided by the manufacturer 2

25.08.2014

3 / 14


Table 2. Chemical composition of blue lupines seeds compared with soybean meal (g/kg as-fed basis) Chemische Zusammensetzung der Samen der Blauen Lupine im Vergleich zu Sojaschrot (g/kg Frischsubstanz) Component Dry matter (DM), g/kg DM composition, g/kg Crude protein Ash Ether extract NDF ADF RFOs Amino acid composition, g/kg CP Arg

Soybean meal

Lupine seeds

909

904

467 64.7

312 34.8

28.5 96.1 64.1 59.1

57.2 254 227 79.8

71.4

91.0

His Ile Leu Lys Met

26.0 45.3 75.7 60.3 13.2

29.8 36.7 64.2 45.8 4.20

Cys Phe Thr Trp

14.5 50.3 39.1 12.8

13.4 38.2 34.9 –

Alkaloids, mg/kg

–

3801

1

angustifoline – 10.7%, isolupanine – 4.63%, lupanine – 62.4%, 13OH lupanine – 22.3%

Sample collection and measurements Feed ingredients and diet samples were analyzed in duplicate for dry matter (DM), crude protein (CP), fat and ash using Association of Official Analytic Chemists (2005) methods 934.01, 976.05, 920.39, and 942.05, respectively. For chemical analysis, the samples were ground to pass through a 0.5 mm sieve. The content of neutral detergent fiber (NDF; assayed with heat stable amylase, aNDF) and acid detergent fiber (ADF), both expressed including residual ash, were determined using the Tecator Foss Fibertec System M (Foss, Hillerod, Denmark). An analysis of raffinose family oligosaccharides (RFOs) was performed using a gas chromatograph 14A (Shimadzu, Kyoto, Japan) equipped with pump LC10ADVP, detector RID 10A and system controller SCL10VP. Amino acids were analyzed using the AAA-339 Mikrotechnika Amino Acid Analyzer, after initial hydrolysis in 6N HCl at 105°C for 23 hours. Before the analysis of sulfur amino acids, performic acid oxidation treatment was carried out to prevent cystine destruction. Alkaloid content was determined at the Plant Variety Testing Laboratory in Slupia Wielka. Alkaloids were extracted from lupine flour with trichloroacetic acid followed by methylene chloride, using a gas chromatograph (Shimadzu GC17A, Kyoto, Japan) with a capillary column (Phenomenex, Torrance, CA), as described by MUZQUIZ et al. (1996). Egg production was monitored for 20 weeks, from 18 to 38 weeks of hen’s age. The body weights of hens were determined at the beginning and at the end of the experiment. Eggs were collected three times a week and egg weight was determined at four-week intervals, for all collected eggs. Feed conversion (kg feed/kg eggs laid) was estimated based on egg weight and feed intake. th

th

th

Towards the end of the 8 , 16 and 20 week of the experiment (at 26, 34 and 38 weeks of hen’s age, respectively), 15 eggs from each group were picked randomly to determine their physicochemical properties. The eggs were weighed, average eggshell thickness was measured using the ESTG-1 eggshell thickness gauge (Orka Food Technology). Next the eggs were broken to assess albumen quality using an egg analyzer (Orka Food Technology), based on albumen height and egg weight expressed in Haugh units. The Haugh unit (HU) was calculated from the 0.37

following formula: HU = log10 (H – 1.7 W

+ 7.56), where H is albumen height (mm), and W is egg weight (g).

Eggshell breaking strength was measured using and egg force reader (Orka Food Technology).

25.08.2014

4 / 14


The concentrations of fatty acids in the yolk were determined in fresh eggs. Fat was extracted from samples by the 3

3

Soxhlet method. Four or five drops (50 – 60 mg) of extracted fat were transferred to 2 cm ampoules, 1.5 cm of a methanol-chloroform-concentrated sulfuric acid (100:100:1 v/v) mixture was added, and the ampoules were sealed

by melting their tops with an open flame. Sealed vials were placed in a dryer at 80–90°C, and were heated for 2 – 3 h. During heating, the ampoules were shaken every 15 min. After solvent evaporation and ampoule cooling, 0.5 – 1 ml of hexane was added to each sample and thoroughly mixed. The resulting fatty acid methyl esters (FAMEs) were

analyzed using a Varian CP-3800 gas chromatograph (Varian, USA) with flame-ionization detection (FID). FAMEs were injected at 225°C, separated on a capillary column (50 m, 0.25 mm, 0.25 μm) (Varian, USA) using a

temperature gradient program (3 min at 50°C, from 50° to 200°C in 40 min) and detected via flame ionization at 250° C. The peaks of fatty acids were identified by comparing their relative retention times with those of individual FAME reference standards (Supelco) diluted in hexane (1:1, 1:2, 1:3, 1:4 v/v). The percentage content of fatty acids and the total pool of PUFAs, including n-3 PUFAs (C18:3n-3, C20:5n-3 C22:5n-3 and C22:6n-3) and n-6 PUFAs (C18:2n-6, C20:2n-6, C20:4 n-6), were determined by relative fatty acid quantification, and % of the total fatty acid peak area was calculated. In week 20 of the laying season, 12 eggs were randomly selected from each group for sensory analysis. A six-member trained panel (ISO 8586–2:1994), experienced in a descriptive analysis of different food products and familiarized with the sensory quality of eggs, performed the assessments. The assessments were carried out in a sensory laboratory room that fulfilled the requirements of the ISO standard (ISO 8589:1998), in individual booths equipped with a computerized system for data collection and processing (FIZZ, Biosystemes, Counternon, France), as described elsewhere (HORSZWALD et al., 2009). For a sensory evaluation, eggs were cooked in boiling water for 12 min, cooled to an external temperature of ca. 20° C, shelled and cut in half. The samples were presented in random order to the assessors from three-digital coded transparent plastic containers covered by lids. Together with the samples, the panelists received a cup of room temperature spring water for cleaning their palates. A quantitative descriptive analysis (QDA) was used to determine the sensory characteristics of the samples. The panelists, together with the panel leader, established the descriptions of the main sensory attributes of eggs using a standard procedure (ISO/DIS 13299:1998). Nine attributes related to the color, aroma, taste and mouth feel (adhesiveness) of eggs were selected and thoroughly defined for profiling. The panelists evaluated the intensity perceived for each of the attributes on a continuous unstructured graphical scale. The scale was 10 cm long and verbally anchored at each end. The left side of the scale corresponded to the lowest intensity and the right side to the highest intensity of the attribute. The results were automatically converted to numerical values (from 0 to 10 units) by a computer. All samples were evaluated in three replications (three sessions) preceded by an introductory session.

Statistical analysis One-way ANOVA was performed to determine the effect of the inclusion level of lupine in experimental diets on laying performance, average egg weight, fatty acid concentrations in yolk lipids, and the sensory properties of eggs. Two-way ANOVA was performed to determine the effects of lupine level, hen’s age and the interaction of both factors (L × A). The significance of differences between mean values of the analyzed parameters in groups was estimated by Duncan’s multiple range test. Data were processed in the STATISTICA PL 9.0 application. In the Tables, results are presented as mean values with pooled standard errors. Data were checked for normal distribution before the statistical analysis was performed. Differences were considered to be significant at p≤0.05.

Results The experimental diets compared in this study had an identical total protein and metabolizable energy content. Diets L10 and L20 had higher concentrations of NDF, ADF and raffinose family oligosaccharides – RFOs (Table 1). In comparison with soybean meal, blue lupine seeds had a lower total protein content (312 vs. 468 g/kg), several-fold higher concentrations of NDF and ADF, and ca. 35% higher RFO levels (79.8 vs. 59.1 g/kg). The total alkaloid content of blue lupine seeds did not exceed 380 mg/kg (Table 2). The inclusion of blue lupine seeds in the ration decreased daily feed intake; significant differences were observed between the control group and the group fed 10% lupine seeds (P = 0.004, Table 3). One-way ANOVA revealed that

25.08.2014

5 / 14


diet supplementation with blue lupine had no effect on other parameters of laying performance. Hens fed lupine seeds produced more eggs, but the noted differences were statistically non-significant. Table 3. Effect of blue lupine on the performance of laying hens. Einfluss der Blauen Lupine auf die Leistung der Legehennen

Group1

Parameter

C

L10

L20

SEM

P

Initial BW2, kg

1.56

1.55

1.55

0.009

0.832

Final BW2, kg

1.99

1.93

1.88

0.018

0.068

Feed intake, g/day

134a

129b

131ab

0.654

0.004

Number of eggs Egg weight, g/hen

129 7621 2.47

130 7579 2.38

132 7640 2.41

0.616 38.6 0.016

0.078 0.811 0.089

FCR3, kg/kg eggs 1

C – control diet, L10 – 10% of blue lupine in the diet, L20 – 20% of blue lupine in the diet BW –body weight, 3FCR – feed conversion ratio a.b Means in the same column without common superscripts differ significantly at p ≤ 0.05 2

There were no significant differences in egg weight between groups; the lowest egg weight was noted in group L20 (Table 4). In weeks 12 and 16 of the laying season, egg weight differences between group L20 and the other groups (including group C characterized by the highest egg weight) approached significance (P = 0.087 and P = 0.065, respectively). Average egg weight during the experiment ranged from 57.4 g in group L20 to 58.6 g in group C. Table 4. Effect of blue lupine on the average egg weight (g) Einfluss der Blauen Lupine auf das durchschnittliche Eigewicht (g)

Group1

C

L10

L20

SEM

P

4 weeks 8 weeks 12 weeks 16 weeks 20 weeks

52.1 57.2 60.6 61.8 62.0

52.2 56.9 59.3 60.2 61.6

51.4 56.4 58.9 60.1 61.0

0.270 0.294 0.323 0.332 0.285

0.395 0.526 0.087 0.065 0.380

Mean

58.6

58.0

57.4

0.256

0.162

1

C – control diet, L10 – 10% of blue lupine in the diet, L20 – 20% of blue lupine in the diet

Two-way ANOVA showed that blue lupine seeds improved eggshell quality, increasing its thickness (P = 0.001), weight as a percentage of total egg weight (P = 0.002), and breaking strength (P = 0.036) (Table 5). Albumen quality, expressed as albumen height (P = 0.001) and Haugh units (P = 0.001), deteriorated with increasing inclusion levels of blue lupine seeds. Blue lupine had no significant effect on other physicochemical properties of eggs. The shells of eggs laid by hens aged 34 and 38 weeks were characterized by significantly lower quality, compared with the shells of eggs produced by 26-week-old hens. Eggshell thickness (P = 0.005), breaking strength (P = 0.004), and weight as a percentage of total egg weight (P = 0.001) were found to decrease. Albumen quality, including albumen height (P = 0.001) and Haugh unit values (P = 0.001), also deteriorated as the hens aged. Eggs produced by older hens had a

25.08.2014

6 / 14


higher yolk content (P = 0.001). A significant lupine level × hen’s age interaction was noted for albumen height (P = 0.037) and eggshell thickness (P = 0.045). Table 5. Effect of blue lupine and hen’s age (weeks) on the physicochemical properties of eggs. Einfluss der Blauen Lupine und des Hennenalters (Wochen) auf die physikochemischen Eigenschaften der Eier Albumen height, mm

Haugh units

Shell thickness, mm

Shell breaking strength, N

Yolk content, %

Group1 CA26

7.97

89.6

CA34

7.79

87.6

CA38

7.60

L10A26

7.88

L10A34

7.50

L10A38 L20A26 L20A34

Albumen content, %

Shell content, %

0.426

4.56

0.386

4.05

24.3

65.5

10.17

25.7

65.0

9.29

86.6

0.403

89.6

0.422

3.90

25.9

64.9

9.24

4.83

24.0

65.6

10.41

86.3

0.421

4.26

25.2

65.2

9.62

6.79

81.8

0.420

7.37

86.7

0.431

4.53

25.3

64.6

10.04

4.63

23.9

65.8

7.01

82.7

0.422

4.33

10.36

26.1

64.2

9.73

L20A38

7.02

82.7

0.420

4.39

26.4

63.7

9.93

SEM

0.061

0.384

0.002

0.063

0.141

0.236

0.067

0

7.79a

87.9a

0.405b

4.17b

25.3

65.1

9.57b

10

7.39b

85.9b

0.421a

4.54a

24.9

65.1

10.02a

20

7.14c 0.001

84.0c 0.001

0.424a 0.001

4.45ab 0.036

25.5

64.5

0.125

0.137

10.01a 0.002

26

7.74a

88.6a

0.427a

4.68a

24.1b

65.6a

10.31a

34

7.43b

85.5b

0.409b

4.22b

25.7a

64.8b

9.54b

38

c

c

0.414 0.005

b

4.28 0.004

a

25.9 0.001

b

0.001

83.7 0.001

b

64.4 0.002

9.74b 0.001

0.037

0.050

0.045

0.661

0.509

0.414

0.496

Lupine (L)

P Age (A)

7.14

P Lx A

2

a.b

Means in the same column without common superscripts differ significantly at p ≤ 0.05 C – control, L – lupine, A– hen’s age 2 lupine level × hen’s age interaction 1

Dietary protein sources had a significant effect on the fatty acid profile of yolk lipids (Table 6). Regardless of their inclusion levels, blue lupine seeds contributed to a significant increase in the concentrations of C18:2 n-6 (P = 0.001) and C18:3 n-3 (P = 0.001) fatty acids and total PUFAs (P = 0.001) in yolk lipids. Eggs from group L20 hens, fed a diet with 20% lupine seeds, had significantly lower concentrations of saturated fatty acids (SFAs) (P = 0.001), and a higher content of C22:6 n-3 fatty acid (P = 0.004) and total UFAs (P = 0.001). As a result, the yolks of eggs from hens fed diets with blue lupine seeds had a higher n-3 and n-6 PUFA content (P = 0.001). The lowest n-6/n-3 PUFA ratio was noted in the yolks of eggs from hens fed 20% lupine seeds (P = 0.035).

25.08.2014

7 / 14


Table 6. Effect of blue lupine on the fatty acid composition of yolk lipids (%) Einfluss der Blauen Lupine auf das Fettsäuremuster der Dotterlipide (%)

Group1

C 14:0 C 14:1 C 15:0 C 16:0

0.42

0.42

a

0.09

b

b

0.11 0.11

L20

L10

C

a

26.5

b

0.11

a

26.0

0.43

SEM

P

0.008

0.799

b

0.004

0.008

a

0.004

0.001

b

0.207

0.001

b

0.154

0.001

0.017

0.001

0.08 0.14

24.9

C 16:1

3.70

2.94

C 17:0

0.29c

0.33b

0.44a

b

b

a

0.015

0.001

b

0.073

0.042

0.253

0.036

C 17:1 C 18:0 C 18:1

a

0.40

ab

8.08

b

0.38 8.37

a

2.51

0.49 7.95

42.0a

40.5b

41.7a

b

a

a

0.335

0.001

0.60a 0.01 0.22 0.13 2.40

0.018

0.001

0.002 0.006 0.004 0.025

0.444 0.911 0.078 0.499 0.640

C 18:2(n-6)

14.2

C 18:3(n-3)

0.46b

C 20:0 C 20:1 C 20:2(n-6) C 20:4(n-6)

0.01 0.22 0.11 2.45

16.4

0.56a 0.01 0.22 0.14 2.48

16.9

C 20:5(n-3)

0.06

0.06

0.07

0.002

C 22:0

0.03b

0.001

0.10

0.04a 0.10

0.001

C 22:5(n-3)

0.03b 0.10

0.005

0.850

C 22:6(n-3)

0.78b

0.83b

0.93a

0.022

0.004

a

a

b

0.212

0.001

SFAs MUFAs PUFAs UFAs n3

35.4

a

46.4

b

18.2

b

64.6

c

1.40

b

n6

16.8

n6/n3

12.0ab

35.2

33.9

c

45.0

b

0.282

0.001

a

a

0.335

0.001

a

0.212

0.001

a

0.033

0.001

19.0

19.4

a

0.338

0.001

12.2a

11.4b

0.169

0.035

44.2 20.6 64.8 1.56

b

b a

21.1

66.1 1.70

1

C – control diet, L10 – 10% of blue lupine in the diet, L20 – 20% of blue lupine in the diet Means in the same column without common superscripts differ significantly at p ≤ 0.05 SFAs – saturated fatty acids MUFAs – monounsaturated fatty acids PUFAs – polyunsaturated fatty acids UFAs – unsaturated fatty acids a..b

The mean sensory ratings for samples and ANOVA results are presented in Table 7. A sensory analysis revealed that yolk color was most intense (P ≤ 0.05) in eggs from hens fed 20% blue lupine seeds (group L20). Blue lupine seeds added to layer diets had no effect on the taste, aroma and texture of eggs. Regardless of the protein source in diets, positive attributes such as egg-like aroma and egg-like taste dominated in all samples.

25.08.2014

8 / 14


Table 7. Average intensity values of egg sensory attributes Durchschnittliche Intensitätsgrade der sensorischen Eigenschaften der Eier

Group1

C

L10

L20

4.6b

4.0b

6.1a

6.3 3.7

6.1 3.7

6.2 3.8

Sulfur-like

0.7

0.7

0.6

Taste Egg-like Buttery Sulfur-like Aftertaste

6.4 3.7 0.5 5.0

6.3 3.9 0.7 5.0.

6.1 4.3 0.6 5.0

Texture Adhesiveness

5.5

5.2

5.6

Attribute Appearance Yolk color Aroma Egg-like Buttery

1

C – control diet, L10 – 10% of blue lupine in the diet, L20 – 20% of blue lupine in the diet Means in the same column without common superscripts differ significantly at p ≤ 0.05 a.b

Discussion According to SWIECICKI and SWIECICKI (1995), the alkaloid content of blue lupine seeds ranges from 0.4 to 3.0% in bitter varieties, but it can be as low as 0.01% in sweet varieties. The total alkaloid content of blue lupine seeds used in our study did not exceed 0.04%, which indicates that Sonet is a low-alkaloid cultivar. Feed intake was significantly lower in hens fed a diet containing 10% lupine seeds, compared with the control group. However, a diet with a higher lupine content did not decrease feed intake, which suggests that lupine-based diets were not less palatable. Experiments performed on pigs have revealed that alkaloids present in lupine-based diets usually reduce feed palatability due to their bitter taste, and thus negatively affect feed intake (KIM et al., 2007; KASPROWICZ-POTOCKA et al., 2013). In the current experiment, lupine seeds used as an alternative protein source had no influence on laying performance. The results of studies investigating the effect of lupine seeds on egg production are inconclusive. LAUDADIO and TUFARELLI (2011) reported that the inclusion of Lupinus albus at 18% in layer diets significantly decreased feed intake, but it had no negative effects on feed efficiency compared with a soybean meal-based diet, which corroborates our findings. Other authors (CUBILLOS et al., 1999; PEREZ-MALDONADO et al., 1999) also noted that the addition of lupine seeds to the diet contributed to high egg production and did not affect other productive parameters. WATKINS and MIROSH (1987) found that egg production in hens fed diets with 30% white lupine was depressed compared with hens fed lower levels of lupine. HAMMERSHØJ and STEENFELDT (2005) demonstrated that blue lupine at different inclusion levels had no effect on feed intake in laying hens, but the feed conversion ratio was negatively correlated with lupine levels. Eggshell breaking strength is a key indicator of egg quality. Previous research (RODRIGEZ-NAVARRO et al., 2002; KRAWCZYK, 2009b) has shown that eggshell breaking strength decreases with hen’s age, most probably due to lower availability of dietary calcium and phosphorus to layers, and changes in shell structure. Also in our study, eggshell breaking strength and other parameters of eggshell quality were lower in older hens. Lupine seeds contained in layer diets improved eggshell quality. In a study by PARK and PARK (2012), eggshell thickness and breaking strength were significantly higher in eggs from hens fed diets containing inulin oligosaccharides. According to some authors (MITSUOKA, 1990; MARTÍNEZ-VILLALUENGA et al., 2008), the use of oligosaccharides, present also in lupine seeds, as natural prebiotics stimulates the proliferation of bifidobacteria in the colon, and the synergistic action of multiple

25.08.2014

9 / 14


bifidobacteria and short-chain fatty acids increases the absorption rate of minerals such as calcium, the main component of eggshells, which was also observed in our study. Other authors (HUGHES and KOCHER, 1998; LAUDADIO and TUFARELLI, 2011) demonstrated that lupine had no effect on eggshell quality.

The differences in albumen height and Haugh unit values, noted in our study, suggest that the replacement of

soybean meal with blue lupine in hen diets deteriorated albumen quality. Our results are consistent with the findings of other authors (HAMMERSHØJ and STEENFELDT, 2005) who demonstrated that albumen dry matter content decreased with increasing lupine levels in layer diets. A decrease in albumen height and, consequently, in Haugh unit values,

observed in our study in hens aged 34 and 38 weeks, was also reported by SILVERSIDES and SCOTT (2001), VAN DEN BRAND et al. (2004) and KRAWCZYK (2009a). The cited authors found that albumen quality deteriorated as the hens aged. According to the USDA (2000), an egg with a Haugh unit score of 72 or more is considered AA grade. Thus, all eggs analyzed in our study were characterized by high quality, irrespective of the diet and age of hens.

In the present experiment, the inclusion of blue lupine seeds in layer diets contributed to an increase in the

concentrations of C18:2 n-6 and C18:3 n-3 fatty acids in yolk lipids. The higher (20%) inclusion rate of lupine seeds in diets enabled to decrease SFA concentrations and improve the n-6/n-3 PUFA ratio in fat extracted from the yolk. There is a scarcity of published research on the effect of lupine seeds on the fatty acid profile of poultry products. UZUN et al. (2007) demonstrated that fatty acids in white lupine seeds were composed of 77% UFAs and 12.6% SFAs, and that the amount of SFAs in lupine was lower than in soybean. Other studies revealed that linoleic acid C18:2 n-6 and α-linolenic acid C18:3 n-3 had the highest share of the total PUFA pool in lupine, which resulted in a very favorable n-6/n-3 PUFA ratio (BOSCHIN et al., 2007, 2008; SUCHÝ et al., 2008). Therefore, the inclusion of blue lupine seeds in layer diets could improve the fatty acid profile of yolk lipids in our study. The QDA used in the present experiment for a sensory evaluation of eggs is a sophisticated tool employed to study a variety of food products (STONE and SIDEL, 1993; LAWLESS and HEYMANN, 1999). No significant differences in the

sensory properties of eggs, including aroma, taste and texture, were observed between groups. HAMMERSHØJ and STEENFELDT (2005) also reported that lupine-based diets had no effect on the sensory properties of eggs, but in another study (VOGT et al., 1983), lupine seeds at 16% resulted in a bitter taste in eggs, whereas the 8% inclusion rate

of lupine had no effect on their sensory properties. In our experiment, a sensory analysis revealed that diets with a 20% lupine content significantly improved yolk color. Other research findings (IGBASAN and GUENTER, 1997; DVORÁK

et al., 2007) also suggest that lupine contained in layer diets contributes to a richer color of egg yolks, most probably due to high concentrations of natural pigments in lupine seeds. PRINSLOO et al. (1992) and QUARANTELLI (1993) did not confirm the beneficial influence of lupine on yolk color. The above differences could be due to the fact that the tested layer diets contained seeds of different lupine varieties with different levels of pigment.

Conclusions Our results indicate that partial replacement of soybean meal (10% or 20%) with blue lupine seeds of cv. Sonet characterized by a low alkaloid content contributed to an increase in the content of NDF, ADF and RFOs, without compromising laying performance. Blue lupine seeds added to layer diets at 20% had a beneficial influence on egg quality – they improved shell quality, yolk color and the fatty acid profile of yolk lipids.

Acknowledgements The study was conducted as part of a long-term program of the Polish Ministry of Agriculture and Rural Development, entitled “Improvements in locally grown vegetable protein sources, vegetable protein production and trading systems and the use of vegetable protein sources in animal feeds”.

25.08.2014

10 / 14


Summary This study was undertaken to determine the effect of partial replacement of soybean meal with 10% and 20% of blue lupine (Lupinus angustifolius) seeds in layer diets on laying performance, the fatty acid profile, physicochemical and

sensory properties of eggs. A 20-week study was performed on 150 Lohman Brown hens with 50 birds in each group. Diet supplementation with blue lupine seeds contributed to an increase in the content of neutral detergent fiber (NDF), acid detergent fiber (ADF) and raffinose family oligosaccharides (RFOs) without compromising laying

performance. Feed intake was lower (P = 0.004) in groups fed blue lupine seeds. Eggshell weight as a percentage of total egg weight, and eggshell breaking strength were higher in groups fed lupine-supplemented diets (P = 0.002 and P = 0.036, respectively). Eggs from hens fed blue lupine seeds were characterized by lower albumen quality expressed

as albumen height (P = 0.001) and Haugh units (P = 0.001). Diet supplementation with blue lupine seeds contributed to an increase in the concentrations of C18:2 n-6 (P = 0.001) and C18:3 n-3 (P = 0.001) fatty acids and total

polyunsaturated fatty acids (PUFAs) (P = 0.001) in yolk lipids. Higher dietary inclusion levels of blue lupine increased the levels of C22:6 n-3 fatty acid (P = 0.004) in the egg yolk, thus significantly (P = 0.035) improving the n-6/n-3 PUFA ratio. Dietary treatments had no influence on the sensory properties of eggs, except for yolk color which was most intense (P ≤ 0.05) in eggs from hens fed 20% blue lupine seeds. Our results indicate that blue lupine seeds can be added to layer diets at 20% without compromising laying performance. Blue lupine seeds improved yolk color and the fatty acid profile of yolk lipids.

Key words Laying hen, nutrition, blue lupine, egg quality, yolk color

Zusammenfassung Fettsäuremuster, physikochemische und sensorische Eigenschaften von Eiern nach Fütterung der Legehennen mit Rationen mit blauen Lupinen Das Ziel der Studie war die Untersuchung des Einflusses eines teilweisen Ersatzes von Sojaextraktionsschrot im Legehennenfutter durch 10 bzw. 20% Samen der Blauen Lupine (Lupinus angustifolius) auf die Leistung, auf das Fettsäuremuster des Dotters und auf physikochemische sowie sensorische Eigenschaften der Eier. Hierzu wurden insgesamt 150 Legehennen auf die drei Behandlungsgruppen verteilt. Der Zusatz der Blauen Lupinen zum Futter erhöhte den Gehalt an neutralen Detergentienfasern (NDF), an sauren Detergentienfasern (ADF) und an Polysacchariden der Raffinose-Familie (RFOs), ohne die Legeleistung zu beeinflussen. Die Futteraufnahme war in den Behandlungen mit Blauen Lupinen geringer (P = 0,004). Das Eischalengewicht bezogen auf das Gesamteigewicht und die Bruchkraft der Eischale waren bei Fütterung mit Blauen Lupinen signifikant höher (P = 0,003 und P = 0,036). Bei Fütterung der Rationen mit Blauen Lupinen war die Eiklarqualität, beschrieben durch die Eiklarhöhe (P = 0,001) und die Haugh-Einheit (P = 0,001), schlechter. Beim Einsatz von Blauen Lupinen nahmen die Gehalte an C18:2 n-6 (P = 0,001) und C18:3 n-3 (P = 0,001) sowie insgesamt an mehrfach ungesättigten (PUFA) Fettsäuren im Dotter signifikant zu. Die höhere Zulage an Blauen Lupinen zum Futter führte zu höheren Gehalten an C22:6 n-3 (P = 0,001) im Dotter und verbesserte so das n-6/n-3 Verhältnis signifikant (P = 0,035). Demgegenüber konnte kein Einfluss auf die sensorischen Eigenschaften der Eier verzeichnet werden. Allerdings war die Dotterfarbe bei der Zulage von 20% Blauen Lupinen am intensivsten (P ≤ 0,05). Die Ergebnisse zeigen, dass Blaue Lupine bis zu einem Anteil von 20% zum Legehennenfutter zugesetzt werden können, ohne negativen Einfluss auf die Leistung. Die Blauen Lupine verbesserten die Farbe und das Fettsäuremuster des Dotters.

Stichworte Legehennen, Fütterung, Blaue Lupine, Eiqualität, Dotterfarbe

References AOAC (Association of Official Analytical Chemists), 2005: Official methods of analysis. 18th ed. Arlington (VA): AOAC.

25.08.2014

11 / 14


BAKER, D.H., 2000: Nutritional constraints to use of soy products by animals. Soy in Animal Nutrition; J.K. Drackley, Ed.; Federation of Animal Science Societies: Savoy, IL, 2000, 1-12. BOSCHIN, G., A. D'AGOSTINA, P. ANNICCHIARICO, A. ARNOLDI, 2007: The fatty acid composition of the oil from Lupinusalbus cv. Luxe as affected by environmental and agricultural factors. Eur. Food Res. Technol. 225, 769776.

BOSCHIN, G., A. D'AGOSTINA, P. ANNICCHIARICO, A. ARNOLDI, 2008: Effect of genotype and environment on fatty acid composition of Lupinus albus L. seed. Food Chem. 108, 600-606. CUBILLOS, A., D. VON BAER, P. GÄDICKE, 1999: Isabrown laying hens fed Lupinus albus or L. angustifolius seed. Lupin, an ancient crop for the new millennium: Proceedings of the 9th International Lupin Conference, Klink/Muritz, Germany, 20-24 June, 1999.2000, 409-410.

DVORÁK, P., E. STRAKOVÁ, J. KUNOVÁ, V. KUNOVÁ, 2007: Egg Yolk Colour Depends upon the Composition of the Feeding Mixture for Laying Hens. Acta Vet. Brno 76, 121-127. HAMMERSHØJ, M., S. STEENFELDT, 2005: Effects of blue lupin(Lupinus angustifolius) in organic layer diets and supplementation with foraging material on egg production and some egg quality parameters. Poultry Sci. 84, 723733. HORSZWALD, A., A. TROSZYŃSKA, M.D. DEL CASTILLO, H. ZIELIŃSKI, 2009: Protein profile and sensorial properties of rye breads. Eur. Food Res. Technol. 229 (6), 875-886. HUGHES, R.J., A. KOCHER, 1998: Nutritive value of lupins for layers. Proceedings of Australian Poultry Science Symposium, Sydney, University of Sydney, 140-143. IGBASAN, F.A., W. GUENTER, 1997: The influence of micronization, dehulling and enzyme supplementation on the nutritional value of peas for laying hens. Poultry Sci. 76, 331-337. JANKOWSKI, J., A. LECEWICZ, Z. ZDUŃCZYK, J. JUŚKIEWICZ, B. SLOMIŃSKI, 2011: The effect of partial replacement of soybean meal with sunflower meal on intestinal adaptation, nutrient utilisation and growth performance of young turkey. Brit. Poultry Sci. 52, 456-465. KASPROWICZ-POTOCKA, M., K. CHILOMER, A. ZAWORSKA, W. NOWAK, A. FRANKIEWICZ, 2013: The effect of feeding Raw and germinated Lupinus luteus and Lupinus angustifolius seeds on the growth performance of young pigs. J. Anim. Feed Sci. 22, 116-121. KIM, J.C., J.R. PLUSKE, B.P. MULLAN, 2007: Lupins as a protein source in pig diets. CAB Reviews: Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources. CAB International, Wallingford 2(3), 1-12. KRAWCZYK, J., 2009a: Effect of layer age and egg production level on changes in quality traits of eggs from hens of conservation breeds and commercial hybrids. Ann. Anim. Sci. 9(2), 185-193. KRAWCZYK, J., 2009b: Quality of eggs from Polish native Greenleg Partridge chicken-hens maintained in organic vs. backyard production systems. Anim. Sci. Pap. Rep. 27(3), 227-235. LAUDADIO, V., V. TUFARELLI, 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. Poultry Sci. 89, 1537-1543. LAUDADIO, V., V. TUFARELLI, 2011: Influence of substituting dietary soybean meal for dehulled-micronized lupin (Lupinus albuscv. Multitalia) on early phase laying hens production and egg quality. Livest. Sci. 140, 184-188. LAWLESS, H.T., H. HEYMANN, 1999: Sensory evaluation of food: principles and practices. Ed., Chapman and Hall, New York, 341-372. MARTÍNEZ-VILLALUENGA, M., J. FRÍAS, C. VIDAL-VALVERDE, 2006: Functional lupin seeds (Lupinus albus L. and Lupinus luteus L.) after extraction of α-galactosides. Food Chem. 98, 291-299.

25.08.2014

12 / 14


MARTÍNEZ-VILLALUENGA, M., C. CHICHOLOSKA, J. KLIBER, A.K. GULEWICZ, 2008: Raffinose family oligosaccharides of st

lupin (LupinusalbusL. cv multolupa) as a potential prebiotic. In: Proceedings of the Nutrition Society, 1 International Immunonutrition Workshop, Valencia, Spain, 67.

MIKULSKI, D., J. JANKOWSKI, Z. ZDUŃCZYK, J. JUŚKIEWICZ, B. SLOMINSKI, 2012: The effect of different dietary levels of rapeseed meal on growth performance, carcass traits, and meat quality of turkeys. Poultry Sci. 91, 215-223. MITSUOKA, T., 1990: Bifidobacteria and their role in human health. J. Ind. Microbiol. 6, 263-268. MUZQUIZ, M., L.M. ROBREDO, C. BURBANO, C. CUADRADO, G. AYET, P. MENDEZ, 1996: Variation In the alkaloid kontent of different subspecies of Chamaecytisus proliferus from the Canary Islands. J. Chromatogr. A. 719, 237-243.

OLKOWSKI, B.I., H.L. CLASSEN, C. WOJNAROWICZ, A.A. OLKOWSKI, 2005: Feeding high levels of lupine seeds to broiler chickens: plasma micronutrient status in the context of digesta viscosity and morphometric and ultrastructural changes in the gastrointestinal tract. Poultry Sci. 84, 1707-1715. PARK, S.O., B.S. PARK, 2012: Effect of feeding inulin oligosaccharides on cecum bacteria, egg quality and egg production in laying hens. Afr. J. Biotechnol. 11(39), 9516-9521. PEREZ-MALDONADO, R.A., P.F. MANNION, D.J. FARREL, 1999: Optimum inclusion of field peas, faba beans, chick peas and sweet lupins in poultry diets. I. Chemical composition and layer experiments. Br. Poultry Sci. 40, 667-673. PETTERSON, D.S., 2000: The use of lupins in feeding systems. Review. Asian Australas. J. Anim. Sci. 13, 861-882. PRINSLOO, J.J., G.A. SMITH, W. RODE, 1992: Sweet white Lupinus albus (cv Buttercup) as a feedstuff for layers. Brit. Poultry Sci. 33, 525-530. PRUSIŃSKI, J., 2007: Biological progress in lupin (Lupinus sp.) – present state and historical background. Zesz. Probl. Post. Nauk Rol. 522, 23-37.

QUARANTELLI, A., 1993: Composition of white lupin seed meal (Lupinu salbus, L.) and its use in feeding laying hens. th

In: Proceedings of the 10

505.

International congress. Scientific Association of Animal Production, Bologna, 499-

RESTA, D., G. BOSCHIN, A. D'AGOSTINA, A. ARNOLDI, 2008: Evaluation of total quinolizidine alkaloids content in lupin flours, lupin-based ingredients, and foods. Mol. Nutr. Food Res. 52, 490-495. RODRIGEZ-NAVARRO, A., O. KALIN, Y. NYS, J.M. GARCIA-RUIZ, 2002: Influence of the microstructure on the shell strength of eggs laid by hens of different ages. Brit. Poultry Sci. 5, 390-415. RUBIO, L.A., A. BRENES, C. CENTENO, 2003: Effects of feeding growing broiler chickens with practical diets containing sweet lupin (Lupinus angustifolius) seed meal. Brit. Poultry Sci. 44, 391-397.

SILVERSIDES, F.G., T.A. SCOTT, 2001: Effect of storage and layer age on quality of eggs from two lines of hens. Poultry Sci. 80, 1240-1245. SMULIKOWSKA, S., A. RUTKOWSKI, 2005: Recommended Allowances and Nutritive Value of Feedstuffs. Poultry Feeding Standards, 4th ed, The Kielanowski Institute of Animal Physiology and Nutrition of the PAS, Jabłonna, Poland (in Polish). StatSoft Inc., 2009: Statistica (data analysis software system) Version 9,0. www.statsoft.com. STRAKOVÁ, E., P. SUCHÝ, I. HERZIG, P. HUDEČKOVÁ, S. IVANKO, 2010: Variation in fatty acids in chicken meat as a result of a lupin-containing diet. Czech J. Anim. Sci. 55, 75-82. STONE, H., J.L. SIDEL, 1993: Sensory evaluation practice. 2nd Ed. Academic Press, San Diego, California. SUCHÝ, P., E. STRAKOVÁ, V. VEČEREK, V. ŠERMAN, N. MAS, 2006: Testing of two varieties of lupin seeds as substitutes for soya extracted meal in vegetable diets designed for young broilers. Acta Vet. Brno. 75, 495-500.

25.08.2014

13 / 14


SUCHÝ, P., E. STRAKOVÁ, L. KROUPA, V. VEČEREK, 2008: The fatty acid content of oil from seeds of some lupin th

varieties. ‘Lupins for Health and Wealth’ Proceedings of the 12 International Lupin Conference, 14-18 Sept. 2008, Fremantle, Western Australia. International Lupin Association, Canterbury, New Zealand, 188-191. SWIECICKI, W., W.K. SWIECICKI, 1995: Domestication and breeding improvement of narrow-leafed lupin (Lupinus angustifolius L.). J. Appl. Genetics 36, 155-167. US Department of Agriculture, 2000: Egg-Grading Manual. Agriculture Marketing Division. Agriculture Handbook No. 75. U.S. Department of Agriculture, Washington, D.C.

UZUN, B., C. ARSLAN, M. KARHAN, C. TOKER, 2007: Fat and fatty acids of white lupin (Lupinus albus L.) in comparison to sesame (Sesamum indicum L.). Food Chem. 102, 45-49. VAN DEN BRAND, H., H. PARMENTIER, K. KEMP, 2004: Effect of housing system (outdoor vs cages) and age of laying hens on egg characteristics. Brit. Poultry Sci. 45 (6), 745-752. VOGT, H., S. HARNISH, R. KRIEG, W. RAUCH, E.C. NABER, 1983: Partly debittered defatted lupin meal in feeds for laying hens. Landbauforsch. Volkenrode 33, 27-30. WIATR, K., A. DOLATA, T. MAŃCZAK, 2007: Lupin cultivars registered In Poland – the concentration and variability of qualitative seed characters. Zesz. Probl. Post. Nauk Rol. 522, 75-85. WATKINS, B.A., L.W. MIROSH, 1987: White lupin as a protein source for layers. Poultry Sci. 66, 1798-1806. ZDUŃCZYK, Z., J. JANKOWSKI, 2013: Poultry meat as functional food: modification of the fatty acid profile – a review. Ann. Anim. Sci. 13, 463-480.

Correspondence: Dr. Aleksandra Drażbo, University of Warmia and Mazury in Olsztyn, Oczapowskiego 5, PL-10718 Olsztyn-Kortowo, Poland; e-mail: [email protected]

25.08.2014

14 / 14