EPS - 10 1399 Eps 2018 227 - Selim 2

See discussions, stats, and author profiles for this publication at: https://www.researchgate.
net/publication/323996086
Effect of Spirulina platensis as a feed additive on laying performance, egg

quality and hepatoprotective activity of laying hens Einﬂuss von Spirulina
platensis als Futterzusatzst...
Article · February 2018
CITATIONS READS
14 1,947
3 authors:
Shaimaa Selim Eman Hussein

Menoufia University Menoufia University
32 PUBLICATIONS 405 CITATIONS 11 PUBLICATIONS 76 CITATIONS
SEE PROFILE SEE PROFILE
Reham Abou-elkhair
Faculty of veterinary medicine, university of Sadat City
20 PUBLICATIONS 219 CITATIONS
SEE PROFILE
Some of the authors of this publication are also working on these related projects:
Effect of supplementing layer hen diet with phytogenic feed additives on laying performance, egg quality, egg lipid peroxidation and blood biochemical constituents
View project
Production performance, egg quality, blood biochemical constituents, egg yolk lipid profile and lipid peroxidation of laying hens fed sugar beet pulp View project
All content following this page was uploaded by Reham Abou-elkhair on 25 March 2018.
The user has requested enhancement of the downloaded file.

Europ.Poult.Sci., 82. 2018, ISSN 1612-9199, © Verlag Eugen Ulmer, Stuttgart. DOI: 10.1399/eps.2018.227
Effect of Spirulina platensis as a feed additive on laying performance, egg

quality and hepatoprotective activity of laying hens
Einfluss von Spirulina platensis als Futterzusatzstoff auf die Legeleistung, die Eiqualität und
den Leberschutz von Legehennen
1* 2 3
S. Selim , E. Hussein and R. Abou-Elkhair
1
Dept. of Nutrition and Clinical Nutrition, Faculty of Veterinary Medicine, University of Menoufia, Egypt
2
Dept. of Poultry Production, Faculty of Agriculture, University of Menoufia, Egypt
3
Dept. of Nutrition and Clinical Nutrition, Faculty of Veterinary Medicine, University of Sadat City, Egypt
*Correspondence: shimaaselim@vet.menofia.edu.eg
Manuscript received 16 November 2017, accepted 15 December 2017
Introduction
The blue-green algae (Spirulina platensis) is widely distributed worldwide. The nutritional value of Spirulina algae is
well-documented. Spirulina has unique high protein content, about 50–70% by dry weight, and it is considered as a
good source of essential amino acids (ANUSUYA et al., 1981). Additionally, Spirulina was recognised to have a wide
range of essential nutrients, including essential fatty acids and polysaccharides, vitamins and minerals, and
carotenoids (DOREAU et al., 2010; GUROY et al., 2012). Some biological activities of Spirulina have been reported.
Spirulina platensis has antioxidant (IBRAHIM and ABDEL-DAIM, 2015), anti-inflammatory (ABDEL-DAIM et al., 2015),
immune-modulating (JAMIL et al., 2015) and hepatoprotective functions (JEYAPRAKASH and CHINNASWAMY, 2005).
Supplementation of Spirulina to animal diets was observed to manage some health problems such as diabetes (GUPTA
et al., 2010), cancer (ISMAIL et al., 2009) and heart diseases (KHAN et al., 2005). The naturally high concentrated
essential nutrients and numerous biochemical and physiological benefits of Spirulina, make it ideal as a natural feed
additive in animal and poultry nutrition, since Spirulina has been found to be safe when added in different food
products (MORAES et al., 2010; BOROWITZKA, 2013).
In poultry diets, algae can be used up to 10% as partial replacement of conventional proteins without any adverse
effects (SPOLAORE et al., 2006). However, levels exceeding 20% resulted in a decline in growth rate of broilers
(TOYOMIZU et al., 2001). Dietary vitamin and mineral premixes can be omitted when Spirulina algae is included in
chicken rations (VENKATARAMAN et al., 1994), due to its nutrients rich composition. Algae could be used as an
effective way to improve the poultry product quality to meet the best consumer preferences (DISMUKES et al., 2008;
SUJATHA and NARAHARI, 2011; ZAHROOJIAN et al., 2013), owing to its high concentration of carotenoids (TAKASHI,
2003; SUJATHA and NARAHARI, 2011). The use of algae as a feed additive, not as a partial protein replacement, in
laying hens is limited. Considering the cost-effectiveness and better egg production and quality, Spirulina could be
used as a valuable natural feed additive. Therefore, the aims of the current study were to determine the effects of
feeding different levels of supplemental algae (Spirulina platensis) on the performance, egg production, egg quality,
blood profiles, and egg yolk cholesterol content of laying hens.
16.02.2018 1 / 13
Materials and Methods

Birds, dietary treatments and management
The experiment was carried out under the protocol approved by the ethics of Faculty of Agriculture, Menoufia
University, Egypt. This study was performed on the Poultry Production Farm, Faculty of Agriculture, Menoufia
University. A total number of 160 Norfa laying hens (a cross of exotic and local egg breeds, White Leghorn × Fayoumi
× White Baladi; KOLSTAD and ABDOU, 2000) at the age of 38 weeks, with an initial body weight of 1057 ± 7.0 g, were
randomly assigned to 4 dietary treatments. There were 4 replicates per treatment with 10 individually caged-
hens/replicate. The cages with a size of 0.44 m high × 0.30 m wide × 0.45 m deep were equipped with nipple drinkers
th th
and trough feeders. The experiment lasted for 8 weeks (from 38 to 46 week of age). Four levels of Spirulina
platensis: 0, 0.1, 0.2 and 0.3% of the diet (0, 1, 2 and 3 kg/ton of the diet, respectively) were used during the
experiment. Pure Spirulina platensis powder (Spirulina ̴ 100%) was obtained from Algal Biotechnology Unit, National
Research Centre, Dokki, Giza, Egypt. The chemical composition of Spirulina platensis was presented in Table 1. The
recommendation tables of NRC (1994) for the nutrient requirements of White-egg layers based on a daily feed intake
of 100 g/hen and egg mass of 50 g/d were utilised to calculate the nutrient requirements for Norfa laying hens (ABD-
EL RAHMAN and ATTIA, 2001; ZANATY, 2005). The composition of the experimental diet was based on NRC (1994)
tabulated values for feedstuffs. The compositions, nutrient concentrations and the proximate chemical analysis,
namely dry matter, crude protein, ether extract, crude fibre and ash (AOAC, 2005) of the basal diet are presented in
Table 2. The hen-house was provided with 16 h light per day. The average temperature in the layer house was
28.6 ± 3.8°C during the experimental periods. Feed and water were provided ad libitum.
16.02.2018 2 / 13
1
Table 1. Nutrient content of Spirulina platensis
Nährstoffgehalt von Spirulina platensis
Items Amount
Chemical composition (% on dry weight basis)

Moisture 6.00
Crude protein 51.7
Crude fat 9.00
Crude fibre 4.50
Ash 10.4
Amino acids (mg/g)

Arginine 48.6
Histidine 16.3
Isoleucine 19.2
Leucine 22.1
Lysine 11.3
Methionine 9.60
Phenylalanine 19.4
Threonine 19.2
Tyrosine 16.7
Valine 14.2
Mineral concentrations (mg/g)

Calcium 83.0
Phosphorus 194
Magnesium 9.20
Iron 16.2
Manganese 1.10
Zinc 3.40
Vitamins/carotenoids (μg/100 g)
Total carotenoids 400–500
Vitamin A 120
Vitamin E 106
Ascorbic acid 540
Fatty acids (%, w/w)

Short chain fatty acids C8:0 1.58
Short chain fatty acids C10:0 6.15
Long chain fatty acids C12:0 1.66
Unsaturated fatty acids 46.7
1
Data were obtained from the manufacturer (Algal Biotechnology Unit, National Research Centre, Dokki, Giza, Egypt).
16.02.2018 3 / 13
Table 2. Composition and nutrient content of the experimental diet
Zusammensetzung und Nährstoffgehalt der Versuchsration
Items Amount
Ingredients, %
Yellow maize 65.8
Soybean meal (44%) 23.7
Vegetable oil 0.65
Sodium chloride 0.34
Dicalcium phosphate1 1.59
Premix 2 0.30
Limestone 7.52
DL-Methionine 3 0.07
Total 100
Calculated energy and nutrient content

ME (MJ/kg) 11.7
Crude protein, g/kg 163
Crude fibre, g/kg 31.1
Ether extract, g/kg 33.4
Calcium, g/kg 32.8
Available phosphorus, g/kg 5.2
Methionine, g/kg 3.4
Lysine, g/kg 9.0
Chemical analysis (g/kg)

DM 902
Crude protein 160
Crude fibre 29.0
Ether extract 32.1
Ash 97.0
1
Dicalcium phosphate, 18% granular phosphate and 23% calcium.
2
Supplied per kg of diet: Vitamin A 12000 IU, vitamin D3 3000 IU, vitamin E 40 mg, vitamin K3 3 mg, vitamin B1 2 mg, vitamin B2 6 mg,
vitamin B6 5 mg, vitamin B12 0.02 mg, niacin 45 mg, biotin 0.075 mg, folic acid 2 mg, pantothenic acid 12 mg, manganese 100 mg, zinc
600 mg, iron 30 mg, copper 10 mg, iodine 1 mg, selenium 0.2 mg, cobalt 0.1 mg. 3DL-Methionine, Met AMINO® (DL-2-amino-4-(methyl-
thio)-butane acid, DL-methionine, α-amino-Y-methyl-oily acid) by Feed Grade 99% (EU).
Sample collection and procedures

th th
Body weights of the laying hens were recorded at the start (38 week of age) and the end of the experiment (46 week
of age). Eggs were collected daily and egg production was calculated on a hen-daily basis. Additionally, egg weight as
well as an egg mass of each treatment group were recorded. Feed intake and egg production were recorded weekly by
th th
replicates to calculate feed conversion ratio (FCR). Eggs produced in the last 3 days of the 4 and 8 week of the
experimental period were individually weighed and analysed for their quality parameters. Egg components,
including albumen, yolk and shell, were weighed separately. Shell thickness was measured by micrometre with a
mean value of measurements at 3 locations on the eggs (near the equator). The height of the albumen and yolk was
determined using a standard tripod micrometre and the diameter of the yolk was measured by calliper. Egg shape
index (%), egg yolk index (%) and egg yolk albumen index (%) were recorded according to ROMANOFF and ROMANOFF
(1949). Egg yolk colour was measured by using the Roche yolk colour fan. Haugh units, a measure of the height of
the albumen of eggs broken out on a flat surface were measured by the use of a tripod micrometre.
th
At the end of the experiment (46 week of age), 8 birds from each treatment (2 hens/replicate) were randomly
selected for blood analysis. Serum levels of cholesterol, ALT and AST were measured by ultraviolet
spectrophotometer using commercially available kits (Biosystem S.A, Costa Brava, 30, Barcelona, Spain) according
to manufacturer’s instructions. In addition, 8 eggs from each treatment group were randomly selected for
determining egg yolk cholesterol content according to the methods described by KAYA et al. (2001) by ultraviolet
spectrophotometer using commercial kits (Biosystem S.A, Costa Brava, 30, Barcelona, Spain).
16.02.2018 4 / 13
Statistical analysis
Experimental data were subjected to One-way ANOVA using IBM SPSS Statistics 22 statistical package (SPSS Inc.,
Chicago, IL, USA) as a completely randomised design. Significant differences among the treatments were determined
using Tukey’s test at P<0.05. In addition, the effect of supplemental levels of Spirulina platensis was determined using
orthogonal polynomials for linear and quadratic effects. Significance was declared as P<0.05 and trends as 0.05≤
P < 0.10. Spearman Rank Correlation Coefficients were calculated using SPSS to identify significant correlations
(P<0.05).
Results
Productive performance
The effects of Spirulina platensis algae as feed additive on the initial body weight (BW), final BW, BW gain and feed
th th th
intake of hens during the period of 38 -46 week of age are presented in Table 3. The initial body weight (38 week
th
old) was not significantly different among the experimental groups. The final BW (46 week old) was significantly
different between the treatment groups (P<0.05). Laying hens fed a diet supplemented with Spirulina platensis algae
at a level of 0.3% of the diet had a higher final BW than the other treatment groups. Also, there was a linear effect on
final BW (P=0.002), as dietary levels of Spirulina platensis increased. Body weight gain was higher in 0.3% Spirulina
platensis group (Control vs. Spirulina, P = 0.08, linear, P=0.02). Data of feed intake indicated a significant difference
among treatments. During the whole 8-week experimental period, hens fed Spirulina platensis algae as supplement
had significantly higher (P<0.01) daily feed intake than those fed the control diet (linear and quadratic, P=0.001).
Table 3. Effect of Spirulina platensis (SP) supplementation on the productive performance of laying hens during the experimental periods
Einfluss des Zusatzes von Spirulina platensis (SP) auf die Leistung der Legehennen in den einzelnen Versuchsperioden
Treatments1 P-value
P-value
Parameters CON T1 T2 T3 SEM2 (CON vs. SP) Linear Quadratic
Body weight, g
Initial BW 1058 1058 1057 1057 7.00 0.996
Final BW 1222c 1227c 1238ab 1253a 7.09 0.011 0.002 0.380
BW Gain 164 170 180 196 11.1 0.081 0.016 0.530
Feed intake, g/hen

Week 38th-42nd 114c 115b 114c 117a 0.136 0.01 0.01 0.01
Week 42 -46 nd th
99 d
110 c
114 a
112 b 0.192 0.001 0.001 0.001
Week 38 -46 th th
107 d
112c
113 b
114 a 0.109 0.001 0.001 0.001
a,b
Values in the same row with a different superscript differ significantly at P ˂ 0.05.
1
CON = control, T1 = 0.1% SP (1 kg/ton), T2 = 0.2% SP (2 kg/ton) and T3 = 0.3% SP (3 kg/ton).
2
Pooled standard error of the means.
16.02.2018 5 / 13
Laying performance
The results of egg number per hen, egg production rate (%), egg weight, egg mass and feed conversion ratio (FCR)
th nd nd th th th
during the period from 38 -42 , 42 -46 and 38 -46 week of age are shown in Table 4. In the first 4 weeks of the
experiment, there were no significant differences in egg number, egg production rate, egg weight, egg mass and FCR
between the treatment groups. However, in the second 4 weeks of the experiment, the inclusion of Spirulina platensis
algae in the diet significantly increased egg number (P<0.001), egg production (P<0.001), and egg mass (P<0.001)
compared to the control group. A tendency was seen towards higher egg weight (P = 0.09) in Spirulina supplemented
groups than in the control group. Laying hens fed a diet supplemented with Spirulina platensis had better feed
th
conversion ratio (P=0.007) during the period 42–46 week of age than the control hens. However, there was no
significant difference in feed conversion ratio during the first 4 weeks of the experiment between the groups.
Spirulina platensis supplemented groups tended to have heavier egg weight (P=0.06) and better FCR (P=0.09) than the
control group during the whole experimental period (38–46 weeks of age). Egg number (linear, P<0.001), egg
production (linear, P<0.001), egg weight (linear, P=0.02) and egg mass (linear, P<0.001) were significantly higher in
th th
laying hens fed 0.3% Spirulina platensis during the experimental period (38 -46 week of age).
Table 4. Effect of Spirulina platensis (SP) supplementation on laying performance during the single experimental periods
Einfluss des Zusatzes von Spirulina platensis (SP) auf die Legeleistung in den einzelnen Versuchsperioden
Treatments2 P-value
P-value
Parameters1 CON T1 T2 T3 SEM3 (CON vs. SP) Linear Quadratic
Egg number, hen

Week 38th-42nd 9.29 9.36 9.49 9.84 0.278 0.254 0.067 0.544
Week 42 -46 nd th
7.51 c
8.47 b
8.88 ab
9.11 a 0.158 < 0.001 < 0.001 0.028
Week 38 -46 th th
8.41 c
8.91 bc
9.19 ab
9.55 a 0.189 0.002 < 0.001 0.591
Egg production, %
Week 38th-42nd 66.5 66.9 67.8 70.3 1.78 0.218 0.060 0.425
Week 42nd-46th 53.6c 60.5b 63.5ab 66.1a 1.13 < 0.001 < 0.001 0.028
Week 38 -46 th th
60.1 c
63.7 bc
65.7 ab
68.2 a 1.35 0.002 < 0.001 0.591
Egg weight, g
Week 38th-42nd 41.9 42.1 42.4 42.5 0.241 0.150 0.036 0.544
Week 42 -46 nd th 45.4 46.1 46.3 46.2 0.346 0.100 0.039 0.141
Week 38 -46 th th 43.6 44.1 44.4 44.4 0.239 0.055 0.016 0.170
Egg mass, g/hen/d

Week 38th-42nd 27.9 28.2 28.8 29.8 0.819 0.167 0.038 0.550
Week 42 -46 nd th
24.3 c
27.9 b
29.4 a
30.5 a 0.481 < 0.001 < 0.001 0.008
Week 38th-46th 26.1c 28.0b 29.1ab 30.2a 0.582 0.001 < 0.001 0.342
Feed conversion ratio

Week 38th-42nd 4.08 4.06 4.00 3.94 0.125 0.683 0.202 0.834
Week 42nd-46th 4.07a 3.92ab 3.87ab 3.75b 0.064 0.007 0.001 0.738
Week 38 -46 th th 4.08 3.99 3.94 3.84 0.079 0.091 0.017 0.977
a,b
1
Egg production = 100 [number of eggs laid ÷ (number of hens × number of days)], Feed conversion ratio = feed intake (g)/egg mass (g).
2
3
16.02.2018 6 / 13
Egg quality measurements

nd th
Data on egg quality parameters and egg yolk score at the end of 42 and 46 week of age are presented in Table 5.
There were no significant differences between the control and Spirulina platensis diets in egg quality traits, including
egg shape index, yolk weight rate, yolk index, albumen weight rate, albumen index, shell weight rate and Haugh
th th
unit at the end of 4 and 8 week of the experiment. However, hens fed Spirulina platensis had a higher shell
nd th
thickness (P<0.001) than those fed the control diet at the end of 42 and 46 week of age. There was linear effect
(P<0.05) on shell thickness as dietary Spirulina platensis increased. Furthermore, the inclusion of Spirulina platensis
resulted in a dose dependent increase in egg yolk colour score (linear and quadratic, P<0.001). There was a strong
positive correlation between the inclusion of Spirulina platensis algae in the diet and egg yolk colour at the end of
nd th
week 42 (r = 0.81, P<0.001) and 46 of age (r = 0.83, P<0.001).
th th
Table 5. Effect of Spirulina platensis (SP) supplementation on egg quality traits at the 4 and 8 week of the experimental period
Einfluss des Zusatzes von Spirulina platensis (SP) auf die Eiqualität am Ende der 4. und der 8. Versuchswoche
Treatments1 P-value
P-value
Parameters Control T1 T2 T3 SEM2 (CON vs. SP) Linear Quadratic
At the end of 42th week of age

Egg Shape Index 74.1 74.2 74.7 76.7 2.27 0.663 0.286 0.593
Shell, % 8.89 8.39 9.12 9.40 0.959 0.941 0.688 0.875
Albumen, % 52.2 53.4 54.9 53.0 1.25 0.253 0.361 0.117
Albumen Index, % 14.7 13.3 12.9 13.1 1.29 0.555 0.241 0.451
Yolk weight, % 38.9 37.2 35.9 37.6 1.16 0.172 0.204 0.078
Yolk shape index, % 44.5 44.8 43.7 44.7 2.31 0.962 0.930 0.846
Yolk colour score 4.78c 6.11b 6.89ab 7.33a 0.272 < 0.001 < 0.001 0.050
Haugh Units, % 90.8 88.3 92.1 89.7 2.32 0.465 0.945 0.985
Shell Thickness, mm 0.314 c
0.356 bc
0.401 ab
0.423 a 0.019 0.002 0.001 0.043
At the end of 46th week of age

Egg Shape Index 75.7 76.3 76.0 74.8 0.954 0.499 0.388 0.222
Shell, % 10.3 9.27 9.84 9.96 0.748 0.582 0.812 0.407
Albumen, % 51.2 54.4 55.4 54.3 2.69 0.469 0.256 0.288
Albumen Index, % 12.1 9.93 10.5 10.7 1.74 0.676 0.561 0.367
Yolk, % 38.5 36.3 34.7 35.7 2.53 0.536 0.253 0.404
Yolk shape index, % 42.7 43.0 40.9 45.0 2.82 0.581 0.627 0.376
Yolk colour score 5.44c 7.44b 7.89ab 8.78a 0.314 < 0.001 < 0.001 0.037
Haugh Units, % 83.6 83.6 87.5 87.9 3.53 0.486 0.172 0.937
Shell Thickness, mm 0.363 b
0.372 ab
0.379 ab
0.405 a 0.021 0.028 0.022 0.134
a,b
1
2
Blood parameters and yolk cholesterol

Results presented in Table 6 show that dietary supplementation with Spirulina platensis significantly decreased
serum cholesterol and ALT concentrations (P<0.001) compared to the control diet. Serum ALT (linear, P<0.001;
quadratic, P=0.004) and cholesterol concentrations (linear, P<0.001) decreased as dietary Spirulina algae increased.
Lower serum level of AST (Control vs. Spirulina, P=0.07, quadratic, P=0.022) in the Spirulina platensis supplemented
groups was observed. Additionally, the lowest significant (linear, P=0.001) total egg yolk cholesterol concentration
was recorded in the group given 0.3%, followed by 0.2% and 0.1% Spirulina platensis. Positive correlation was
observed between the inclusion of Spirulina platensis algae in the diet of laying hens and decreasing both serum
(r = 0.75, P<0.001) and egg yolk (r = 0.78, P<0.001) total cholesterol concentrations at the end of the experiment.
16.02.2018 7 / 13
Table 6. Effect of Spirulina platensis (SP) supplementation on selected serum parameters of laying hens and egg yolk cholesterol
th
concentration at the end of the experiment (46 week of age)
Einfluss des Zusatzes von Spirulina platensis (SP) auf ausgewählte Blutparameter der Legehennen und auf den Dotter-Cholesterol-Gehalt am
Versuchsende (46. Lebenswoche)
Treatments2 P-value
P-value
Parameters1 CON T1 T2 T3 SEM3 (CON vs. SP) Linear Quadratic
Cholesterol, mg/dl 135a 130ab 123b 115c 2.13 < 0.001 < 0.001 0.302
AST, U/l 24.1 24.0 23.9 23.8 0.109 0.073 0.183 0.022
ALT, U/l 12.4a 12.1b 9.91bc 9.82c 0.084 < 0.001 < 0.001 0.004
Yolk cholesterol, mg/g 13.6a
13.1 a
12.4 ab
11.7 b 0.397 0.005 0.001 0.710
a,b
1
AST = Aspartate aminotransferase, ALT = alanine aminotransferase.
2
3
Discussion
Algae can be used as a valuable ingredient in human and animal nutrition because of its unique nutritional profile as
the algae is rich in essential nutrients such as essential amino acids, vitamins, minerals, antioxidants and other
bioactive substances (DOREAU et al., 2010; AL-HARTHI and EL-DEEK, 2011; AL-HARTHI and EL-DEEK, 2012; GUROY et
al., 2012; AL-HARTHI, 2014). In the current study, we aimed to determine the beneficial effects of feeding blue-green
algae (Spirulina platensis) as a feed additive on the laying performance, egg quality traits and blood profile of laying
hens.
Dietary inclusion of Spirulina platensis at a level 0.1, 0.2 and 0.3% in the diets of laying hens increased their body
weight and feed intake during the experiment. Our findings are in agreement with previous studies (NIKODÉMUSZ et
al., 2010; MARIEY et al., 2012), who reported beneficial effects of dietary Spirulina algae supplementation on the
productive performance of poultry. However, HALLE et al. (2009) reported a significant reduction in feed intake of
laying hens when 7.5 g/kg bullet-milled and spray-dried Chlorella vulgaris was included in their diets. The increase in
body weight of hens after 8 weeks feeding Spirulina platensis in the current study may contribute to improving feed
utilisation efficiency (NIKODÉMUSZ et al., 2010; KAOUD, 2012; MARIEY et al., 2012), enhancing the absorption of
vitamins and minerals and to optimising the nutrient digestion process (GRUŽAUSKAS et al., 2004).
Inclusion of Spirulina platensis to the diet of laying hens improved their egg production rate, egg weight, egg mass
and FCR during the whole 8-week experimental period. In fact, this improvement was notable after 4 weeks of
feeding Spirulina algae-supplemented diet. KAOUD (2012) observed that dietary inclusion of Spirulina improved FCR
of broilers. Previous studies (NIKODÉMUSZ et al., 2010; MARIEY et al., 2012) recorded that laying birds fed diets
supplemented with Spirulina algae achieved better egg production rates and FCR than those fed the control diet.
Dietary supplementation of Chlorella has been shown to improve laying performance of layers (ZHENG et al., 2012).
On the contrary, ZAHROOJIAN et al. (2011) observed no differences in egg production, feed intake, FCR and egg weight
when marine algae (Spirulina platensis) was added at a level of 1.5, 2 and 2.5% to the diet of Hy-Line hens.
Furthermore, CARRILLO et al. (2008) studied the effect of different marine algae (Macrocystic pyrifera, Saryassum
sinicola and Enteromorpha sp.) at a level of 10% of each in Leghorn hens’ diets and observed that egg production, feed
intake, FCR and egg weight were not affected by the inclusion of marine algae. It seems that this variation in the
obtained results between the current study and previous ones may be contributed to the inclusion levels, type of
algae, and breed of laying hens. The current study may not be able to completely explain the observed positive
response of feeding Spirulina platensis in laying performance. However, microalgae were reported to contain
beneficial components such as essential amino acids, vitamins, carotenoids, antioxidants, antimicrobial factors and
n-3 fatty acids (ABD EL-BAKY et al., 2008; AL-HARTHI and EL-DEEK, 2011; AL-HARTHI and EL-DEEK, 2012; AL-HARTHI,
2014). Furthermore, the positive effect of feeding Chlorella algae on the caecal microbiota profiles of layers has been
16.02.2018 8 / 13
reported (KIM and KANG, 2015). Thus, these above mentioned factors could be included in the metabolism regulating
egg production of layers.
In the present study, the effects of dietary inclusion of Spirulina algae on the percentages of egg shell, albumen and
yolk, and the egg shape index or the Haugh unit were not significantly different. These findings were in agreement
with previous studies (ZAHROOJIAN et al., 2011; MARIEY et al., 2012). On the other hand, our study observed that the
inclusion of Spirulina platensis increased the egg shell thickness. In line with our finding, PARK et al. (2015) showed
that supplementation of marine microalgae (Schizochytrium) powder at a level 0.5–1% of the diet had a beneficial
effect on the egg shell thickness. The influence of marine algae on shell thickness may be attributed to its mineral
content (PARK et al., 2015). However, the mechanisms underlying this effect still remain unclear, and there was no
scientific evidence explaining how algae affect the egg shell thickness. Further investigations are needed to study the
relation between Spirulina platensis and mineral metabolism of laying hens.
Algae has a natural beneficial impact on human health through improving the quality of eggs by enhancing total
carotenoids and antioxidant content and decreasing yolk cholesterol concentration (ZAHROOJIAN et al., 2013; AL-
HARTHI, 2014; PARK et al., 2015). A dose dependent increase in the egg yolk colour score as a result of addition
Spirulina platensis algae in layer diets of the present study most likely reflects the deposition of carotenoids in yolk
and resulting in increased egg yolk pigmentation. These findings agreed with previous studies (SUJATHA and
NARAHARI, 2011; MARIEY et al., 2012; ZAHROOJIAN et al., 2013; AL-HARTHI, 2014). In this regard, AL-HARTHI (2014)
and KOTRBACEK et al. (2013) indicated that carotenoids are efficiently deposited in the egg yolk, when included in
layer diets.
There was a significant decrease in yolk cholesterol, and serum cholesterol ALT and AST concentrations in groups
fed Spirulina platensis algae. AL-HARTHI and EL-DEEK (2012) reported that supplementation of brown marine algae in
laying hen diets significantly decreased egg yolk cholesterol, and this effect may be due to the antioxidant properties
of marine algae. The lower levels of egg yolk cholesterol were related to their reduced levels in blood of hens fed
Spirulina-containing diets (MARIEY et al., 2012; ZAHROOJIAN et al., 2013). Additionally, the decreased blood
cholesterol observed in Spirulina-supplemented groups could be related to docosahexaenoic acid (PARK et al., 2015)
and dietary fibre in marine algae (LAHAYE and JEGOU, 1993). Dietary fibre or plant sterols were reported to inhibit the
absorption of intestinal cholesterol (LIRETTE et al., 1993). The observed reduction in serum ALT and AST levels of
Spirulina-supplemented groups in the current study revealed the hepatoprotective activity of the Spirulina platensis as
a result of its antioxidant and anti-inflammatory factors.
Conclusions
The results of the current experiment indicate that dietary supplementation with Spirulina platensis improves the
laying performance, egg quality and hepatoprotective activity. As a consequence, dietary Spirulina platensis at a level
of 0.3% of the diet (3 kg/ton) is considered a valuable natural feed additive for laying hens. Further investigations are
needed to study the relation between dietary inclusion of Spirulina platensis algae and mineral metabolism of laying
hens.
Acknowledgements
The authors extend their appreciation to scientific research at the Faculty of Agriculture, University of Menofia and
Central Lab of Blood and Chemical Analysis at the Faculty of Veterinary Medicine, University of Sadat City. The
authors are thankful to Department of Poultry Production, Faculty of Agriculture, Menofia University and
Department of Nutrition and Clinical Nutrition, Faculty of Veterinary Medicine, Sadat City University for providing
the financial support and research facilities.
Conflict of interest declaration

The authors declare that there was no conflict of interest.
16.02.2018 9 / 13
Summary
This study aimed to determine the effect of different dietary levels of Spirulina platensis on the laying performance,
egg quality traits and hepatoprotective status of laying hens. A total of 160 Norfa laying hens (38 weeks old) were
randomly allocated to one of 4 dietary treatments, with 4 replicates per treatment. The basal diet was used and three
additional diets were prepared by supplementing 0.1, 0.2 or 0.3% of Spirulina platensis to the basal diet. The diets
were fed to laying hens ad libitum for 8 weeks. Egg production, feed intake, feed conversion ratio, egg weight, egg
mass, internal and external egg quality parameters were recorded. Egg yolk cholesterol, serum cholesterol, alanine
aminotransferase (ALT) and aspartate aminotransferase (AST) concentrations were measured.
Results indicated that supplementation of Spirulina to the diet of laying hens increased (P<0.05) their body weight
and feed intake compared to the control. Dietary inclusion of Spirulina platensis improved egg number per hen, egg
weight, egg production, egg mass and feed conversion ratio (linear, P<0.05) during the whole experimental period.
No significant differences were observed in the egg shape index and the percentages of egg shell, albumen, yolk or
Haugh unit by the inclusion of Spirulina platensis in diets. However, hens fed Spirulina platensis had a higher shell
thickness (P<0.05) than those fed the control diet. Inclusion of Spirulina platensis resulted in a dose dependent
increase in egg yolk colour score (linear and quadratic, P<0.001). A strong positive correlation was observed between
the inclusion of Spirulina platensis algae and egg yolk colour score. Dietary supplementation of Spirulina platensis
decreased serum cholesterol (linear, P<0.001) and ALT (linear and quadratic, P<0.01) and AST concentration
(quadratic, P=0.02). Egg yolk cholesterol concentration was decreased as the dietary concentration of Spirulina algae
increased (linear, P=0.001). These results suggest that Spirulina platensis at a level of 0.3% can be used as a functional
natural feed additive to improve laying performance, egg quality and hepatoprotective activity.
Key words
Laying hen, nutrition, Spirulina platensis, performance, egg quality, hepatoprotective activity
Zusammenfassung
Einfluss von Spirulina platensis als Futterzusatzstoff auf die Legeleistung, die Eiqualität und den Leberschutz von
Legehennen
Im Rahmen der Studie wurde der Effekt der Zulage von Spirulina platensis in unterschiedlicher Höhe zum Futter auf
die Leistung, die Eiqualität und den Leberschutz bei Legehennen untersucht. Hierzu wurden insgesamt 160 Norfa
Legehennen (38 Wochen alt) zufällig auf 4 Behandlungsgruppen mit je 4 Wiederholungen verteilt. Der gemeinsamen
Basisfutterration, die auch als Kontrollgruppe diente, wurden 0,1, 0,2 oder 0,3% Spirulina platensis zugesetzt. Die
Rationen wurden über 8 Wochen verfüttert und die Legeleistung, die Futteraufnahme, die Futterverwertung, die
tägliche Eimasse, die Gesamteimasse und Parameter der äußeren und der inneren Eiqualität erfasst. Ferner wurden
der Cholesterol-Gehalt im Dotter und im Blutserum sowie die Konzentrationen an Alanin-Amino-Transferase (ALT)
sowie Aspartat-Amino-Transferase (AST) im Blut bestimmt.
Die Zulage von Spirulina platensis hat bei den Legehennen im Vergleich zur Kontrolle zu höheren Tiergewichten und
Futteraufnahme geführt (P < 0,05), die Eizahl je Henne, das Eigewicht, die Legeleistung, die Eimasseproduktion und
die Futterverwertung nahmen über die Versuchsdauer ebenfalls zu (P < 0,05). Dagegen konnten keine Effekte auf die
Eiform, auf den Schalen-, Eiweiß- und Dotteranteil sowie auf die Haugh-Einheit beobachtet werden. Allerdings war
bei den Behandlungsgruppen die Schalendicke signifikant höher als in der Kontrollgruppe (P < 0,05). Mit
zunehmender Zulage an Spirulina platensis nahm die Dotterfarbe signifikant zu (linear und quadratisch, P < 0,001).
Es lag somit eine klare positive Korrelation zwischen der Zulage von Spirulina platensis und der Dotterfarbe vor. Die
Zulage an Spirulina platensis verminderte den Serum-Cholesterol-Spiegel (linear, P < 0,001) und die Konzentrationen
von ALT (linear und quadratisch, P < 0,01) sowie AST (quadratisch, P < 0,02) im Blutserum. In ähnlicher Weise
nahm mit der Zulage an Spirulina platensis der Dotter-Cholesterol-Gehalt ab (linear, P = 0,001). Die Ergebnisse deuten
darauf hin, dass eine Zulage von 0,3% zum Futter die Legeleistung und die Eiqualität positiv beeinflussen und auch
den Leberschutz verbessern kann.
Stichworte
Legehenne, Fütterung, Spirulina platensis, Legeleistung, Eiqualität, Leberschutz
16.02.2018 10 / 13
References
ABD EL-BAKY, H.H., F.K. EL BAZ, G.S.E. BAROTY, 2008: Evaluation of marine alga Ulva Lactuca l. as a source of
natural preservative ingredient. Am-Eurasian. J. Agric. Environ. Sci. 3, 434-444.
ABD-EL RAHMAN, S.A., Y.A. ATTIA, 2001: Response of Norfa white egg breeders to amino acid supplementations to
low protein diets. Arch. Geflugelk. 66, 35-42.
ABDEL-DAIM, M.M., S.M. FAROUK, F.F. MADKOUR, S.S. AZAB, 2015: Anti-inflammatory and immunomodulatory
effects of Spirulina platensis in comparison to Dunaliella Salina in acetic acid-induced rat experimental colitis.
Immunopharm. Immunot. 37, 126-139.
AL-HARTHI, M.A., 2014: The effect of natural and synthetic antioxidant on performance, egg quality and blood
constituents of laying hens grown under high ambient temperature. Ital. J. Anim. Sci. 13. DOI:
10.4081/ijas.2014.3239.
AL-HARTHI, M.A., A.A. EL-DEEK, 2011: The effects of preparing methods and enzyme supplementation on the
utilization of brown marine algae (Sargassum dentifebium) meal in the diet of laying hens. Ital. J. Anim. Sci. 10.
DOI: 10.4081/ijas.2011.e48.
AL-HARTHI, M.A., A.A. EL-DEEK, 2012: Effect of different dietary concentrations of brown marine algae (Sargassum
dentifebium) prepared by different methods on plasma and yolk lipid profiles, yolk total carotene and lutein plus
zeaxanthin of laying hens. Ital. J. Anim. Sci. 11. DOI: 10.4081/ijas.2012.e64.
ANBARASAN, V., V.K. KUMAR, P.S. KUMAR, T. VENKATACHALAM, 2011: In vitro evaluation of antioxidant activity of
blue green algae Spirulina platensis. Int. J. pharm. Sci. Res. 2, 2616-1618.
ANUSUYA, D.M., G. SUBBULAKSHIMI, D.K. MADHAVI, L.V. VENKATARAM, 1981: Studies on the proteins of mass-
cultivated, blue-green alga (Spirulina platensis). J. Agric. Food Chem. 29. DOI: 10.1021/jf00105a022.
AOAC, 2005: Official Methods of Analysis of the Association of Analytical Chemists International, 18th ed.
Gathersburg, MD U.S.A Official methods, 2005.08.
BOROWITZKA, M.A., 2013: High-value products from microalgae-their development and commercialisation. J.
Applied Phycol. 25, 743-75.
CARRILLO, S., E. LO´PEZ, M. CASAS, E. AVILA, R.M. CASTILLO, M.E. CARRANCO, C. CALVO, F. PEREZ-GIL, 2008: Potential
use of seaweeds in the laying hen ration to improve the quality of n-3 fatty acid enriched eggs. J. Appl. Phycol. 20,
271-278.
DISMUKES, G.C., D. CARRIERI, N. BENNETTE, G.M. ANANYEV, M.C. POSEWITZ, 2008: Aquatic phototrophs: efficient
alternatives to land-based crops for biofuels. Curr. Opin. Biotechnol. 19. DOI:org/10.1016/j.copbio.2008.05.007.
DOREAU, M., D. BAUCHART, Y. CHILLIARD, 2010: Enhancing fatty acid composition of milk and meat through animal
feeding. Anim. Prod. Sci. 51, 19-29.
GRUŽAUSKAS, R., R. LEKAVICIUS, R. RACEVICIUT-STUPELIEN, V.T. ŠAŠYT, G.J. ŠVIRMICKAS, 2004: Višiuk broileriu
virškinimo procesu optimizavimas simbiotiniais preparatais. Veterinarija ir Zootechnika 28, 51-56.
GUPTA, S., H.J. HRISHIKESHVAN, P.K. SEHAJPAL, 2010: Spirulina protects against rosiglitazone induced osteoporosis in
insulin resistance rats. Diabetes. Res. Clin. Pract. 87. DOI: 10.1016/j.diabres.2009.10.002.
GUROY, B., I. SAHIN, S. MANTOGLU, S. KAYALI, 2012: Spirulina as a natural carotenoid source on growth,
pigmentation and reproductive performance of yellow tail cichlid Pseudotropheus acei. Aquacult. Int. 20, 869-
878.
HALLE, I., P., JANCZYK, G. FREYER, W.B. SOUFFRANT, 2009: Effect of microalgae Chlorella vulgaris on laying hen
performance. Arch. Zootech. 12, 5-13.
IBRAHIM, A.E., M.M. ABDEL-DAIM, 2015: Modulating effects of Spirulina platensis against tilmicosin-induced
cardiotoxicity in mice. Cell J. 17, 137-144.
16.02.2018 11 / 13
ISMAIL, M.F., D.A. ALI, A. FERNANDO, M.E. ABDRABOH, R.L. GAUR, 2009: Chemoprevention of rat liver toxicity and
carcinogenesis by Spirulina. Int. J. Biol. Sci. 5. DOI: 10.7150/ijbs.5.377.
JAMIL, A.B.M.R., M.R. AKANDA, M.M. RAHMAN, M.A. HOSSAIN, M.S. ISLAM, 2015: Prebiotic competence of Spirulina
on the production performance of broiler chickens. J. Adv. Vet. Anim. Res. 2. DOI: 10.5455/javar.2015.b94.
JEYAPRAKASH, K., P. CHINNASWAMY, 2005: Effect of Spirulina and Liv-52 on cadmium induced toxicity in albino rats.
Indian. J. Exp. Biol. 43, 773-781.
KAOUD, A.H., 2012: Effect of Spirulina platensis as a dietary supplement on broiler performance in comparison with
prebiotics. Scient. J. Appl. Res. 1, 44-48.
KAYA, S., T. KECECI, S. HALILOĞLU, 2001: Effects of zinc and vitamin A supplements on plasma levels of thyroid
hormones, cholesterol, glucose and egg yolk cholesterol of laying hens. Res. Vet. Sci. 71. DOI:
10.1053/rvsc.2001.0500.
KHAN, M., J.C. SHOBHA, I.K. MOHAN, M.U.R. NAIDU, C. SUNDARAM, 2005: Protective effect of Spirulina against
doxorubicin-induced cardiotoxicity. Phytother. Res. 19. DOI: 10.1002/ptr.1783.
KIM, C.H., H.K. KANG, 2015: Effect of dietary supplementation with a Chorella by-product on the performance,
immune response and metabolic function in laying hens. Europ. Poult. Sci. 79. DOI: 10.1399/eps.2015.108.
KOLSTAD, N., F.H. ABDOU, 2000: NORFA: The Norwegian-Egyptian project for improving local breeds of laying hens in
Egypt. Workshop on Developing Breeding Strategies for Lower Input Production Environment, Bella, Italy, In:
Proceedings of the ICAR Technical Series. 3, 301-306.
KOTRBACEK, V., M. SKRIVAN, J. KOPECKY, O. PENKAVA, P. HUDECKOVA, I. UHRÍKOVA, J. DOUBEK, 2013: Retention of
carotenoids in egg yolks of laying hens supplemented with heterotrophic Chlorella. Czech. J. Anim. Sci. 58, 193-
200.
LAHAYE, M., D. JEGOU, 1993: Chemical and physical-chemical characteristics of dietary fibers from Ulva lactuca (L.)
Thuret and Enteromorpha compressa (L.) Grev. J. Appl. Physiol. 5, 195-200.
LIRETTE, A., A.R. ROBINSON, D.C. CROBER, P.D. LAWSON, N.L. FIRTH, 1993: Effect of oat bran, cotton seed hulls and
guar gum on chicken egg and blood lipids during the early laying period. Can. J. Anim. Sci. 73.
doi.org/10.4141/cjas93-074.
MARIEY, Y.A., H.R. SAMAK, M.A. IBRAHEM, 2012: Effect of using Spirulina platensis algae as a feed additive for poultry
diets: 1-Productive and reproductive performances of local laying hens. Egypt. Poult. Sci. 32, 201-215.
MORAES, C.C., J.F. DE MEDEIROS BURKERT, S.J. KALIL, 2010: C-phycocyanin extraction process for large-scale use. J.
Food Biochem. 34, 133-148.
NIKODÉMUSZ, E., P. PÁSKAI, L. TÓTH, J. KOZÁK, 2010: Effect of dietary Spirulina supplementation on the reproductive
performance of farmed pheasants. Tech. Articles-Poult Ind. pp. 1-2.
NRC, 1994: Nutrient Requirements of Poultry. Ninth ed. Natl. Acad. Press. Washington, DC.
PARK, J.H., S.D. UPADHAYA, I.H. KIM, 2015: Effect of dietary marine microalgae (schizochytrium) powder on egg
production, blood lipid profiles, egg quality, and fatty acid composition of egg yolk in layers. Asian-Australasian.
J. Anim. Sci. 28, 391-397.
ROMANOFF, A.L., A.L. ROMANOFF, 1949: The Avian Egg. John Wiley. And Sons, Inc., New York.
SPOLAORE, P., C. JOANNIS-CASSAN, E. DURAN, A. ISAMBERT, 2006: Commercial Applications of Microalgae. J. Biosci.
Bioeng. 101. DOI: org/10.1263/jbb.101.87.
SUJATHA, T., D. NARAHARI, 2011: Effect of designer diets on egg yolk composition of ‘White Leghorn’ hens. J. Food
Sci. Tech. 48, 494-497.
TAKASHI, S., 2003: Effect of administration of Spirulina on egg quality and egg components. Anim. Husbandry. 57,
191-195.
16.02.2018 12 / 13
TOYOMIZU, M., K. SATO, H. TARODA, T. KATO, Y. AKIBA, 2001: Effects of dietary Spirulina on meat colour in muscle of
broiler chickens. Br. Poult. Sci. 42. DOI:10.1080/00071660120048447.
VENKATARAMAN, L.V., T. SOMASEKARAN, E.W. BECKER, 1994: Replacement value of blue-green alga (Spirulina
platensis) for fishmeal and a vitamin-mineral premix for broiler chicks. Br. Poult. Sci. 35. DOI:
org/10.1080/00071669408417702.
ZAHROOJIAN, N., H. MORAVEJ, M. SHIVAZAD, 2011: Comparison of marine algae (Spirulina platensis) and synthetic
pigment in enhancing egg yolk colour of laying hens. Br. Poult. Sci. 52, 584-588.
ZAHROOJIAN, N., H. MORAVEJ, M. SHIVAZAD, 2013: Effects of dietary marine algae (Spirulina platensis) on egg quality
and production performance of laying hens. J. Agr. Sci. Tech. 15, 1353-1360.
ZANATY, G.A, 2005: Optimum dietary protein and energy levels for Norfa hens during the laying period. Egypt.
Poult. Sci. 27, p 207.
ZHENG, L., S.T. OH, J.Y. JEON, B.H. MOON, H.S. KWON, S.U. LIM, B.K. AN, C.W. KANG, 2012: The dietary effects of
fermented Chlorella vulgaris (CBT®) on production performance, liver lipids and intestinal microflora in laying
hens. Asian. Austral. J. Anim. Sci. 25, 261-266.
16.02.2018
View publication stats
13 / 13

EPS - 10 1399 Eps 2018 227 - Selim 2

Uploaded by

Document Information

Original Title

Copyright

Available Formats

Share this document

Share or Embed Document

Sharing Options

Did you find this document useful?

Is this content inappropriate?

Copyright:

Available Formats

EPS - 10 1399 Eps 2018 227 - Selim 2

Uploaded by

Copyright:

Available Formats

See discussions, stats, and author profiles for this publication at: https://www.researchgate.

Effect of Spirulina platensis as a feed additive on laying performance, egg

Article · February 2018

Shaimaa Selim Eman Hussein

SEE PROFILE SEE PROFILE

The user has requested enhancement of the downloaded file.

Effect of Spirulina platensis as a feed additive on laying performance, egg

Manuscript received 16 November 2017, accepted 15 December 2017

Materials and Methods

Nährstoffgehalt von Spirulina platensis

Chemical composition (% on dry weight basis)

Amino acids (mg/g)

Mineral concentrations (mg/g)

Fatty acids (%, w/w)

Table 2. Composition and nutrient content of the experimental diet

Zusammensetzung und Nährstoffgehalt der Versuchsration

Calculated energy and nutrient content

Chemical analysis (g/kg)

Sample collection and procedures

Feed intake, g/hen

Egg number, hen

Egg mass, g/hen/d

Feed conversion ratio

Egg quality measurements

At the end of 42th week of age

At the end of 46th week of age

Blood parameters and yolk cholesterol

Conflict of interest declaration

You might also like