Livestock Research for Rural Development 33 (8) 2021 LRRD Search LRRD Misssion Guide for preparation of papers LRRD Newsletter

Citation of this paper

Effects of dietary inclusion of Spirulina platensis on egg yolk pigmentation

F Khan, A A S Shuvo, M J Khan and K M S Islam

Department of Animal Nutrition, Bangladesh Agricultural University, Bangladesh


The present experiment was conducted to compare the effects of spirulina ( Spirulina platensis) on egg quality and the level of pigmentation in yolk of egg. A total number of 120 Shaver-579 hens were used and the experiment was continued for 7 weeks. The hens were randomly assigned into 4 treatment groups (6 replications and 5 hens per replication) in completely randomized design (CRD). Wheat based diet was considered as basal diet. No spirulina was supplied in control diet (0%) but 0.1, 0.2 and 0.3% spirulina was supplied in other three diets (0.1, 0.2 and 0.3%). Number of eggs laid and weight of egg recorded daily. The internal egg qualities were determined by estimating the albumen weight, albumen index, Haugh Unit, yolk index and yolk color. The external egg quality characteristics were measured by estimating the egg shape index, shell thickness, dry shell weight, percent shell. Egg weight and egg mass output was high in 0.2% diet group (p<0.05). Albumen index and Haugh unit also significantly high in 0.2% spirulina supplemented dietary group. The value for L* and a* were significantly different, L* was high (p<0.05) in 0.2% and a* was high in 0.2% which indicates the higher deposition of carotene in egg. From the result it could be concluded that spirulina helps in carotene deposition in yolk that would enhance the consumer preference for egg.

Keywords: egg quality, microalgae, layer


Market and demand for animal products are expanding in developing countries due to the changes of consumer’s taste. Now a days consumer prefers safe and nutritious animal products. The price of conventional feedstuffs has risen dramatically in the poultry industry, owing to a lack of supplies and food-feed competition. As a result, researchers and producers are both interested in exploring new feed resources for poultry. Microalge has a high production rate and a higher nutritional value for monogastric animals, microalge such as Spirulina platensis could be one of the greatest solutions for better development, production performance, and egg quality. Essential fatty acids and polysaccharides, vitamins and minerals, and carotenoids are all found in spirulina (Doreau et al 2010; Guroy et al 2012). Spirulina has been linked to a variety of biological functions. Spirulina platensis contains antioxidant (Abdel-daim, 2015), anti-inflammatory, immune-modulating (Jamil et al 2015), and hepatoprotective properties. Due to its high concentration of carotenoids (Takashi et al 2003; Sujatha et al 2011), inclusion of algae in poultry diet could be an effective technique to improve the quality of poultry products to suit the best consumer preferences (Dismukes et al 2008; Sujatha et al 2011; Zahroojian et al 2013). Moreover, Takashi (2003) found that egg yolk color was significantly improved by the addition of Spirulina to laying hen diets that might be due to proper pigmentation and high carotene deposition that comes from spirulina. The present experiment was design to evaluate the effect of spirulina on production performance of layer birds, egg quality and egg yolk pigmentation.

Materials and methods

Experimental layout, diet and management practices

The Animal Welfare and Experimental Ethics Committee of Bangladesh Agricultural University (AWEEC/BAU/2020/31) examined and approved the experimental techniques, animal care, and sample collection. 120 Shaver-579 hens were randomly assigned in 4 treatment groups having 6 replications (5 hens per replication) and were reared in three tiered cage for 7 weeks. Wheat based diet (CP=17.9%, ME=2750 Kcal/kg) were offered to the layer birds of all dietary treatment groups. No spirulina was supplied in 0% group but 0.1, 02 and 0.3% spirulina was supplied to dietary treatment groups of 0.1, 02 and 0.3%. Completely wheat based diets were formulated by using locally available raw materials. Different raw ingredients like wheat, soyabean oil, soyabean meal, DCP. were purchased from local market. The ration and calculated composition were shown in Table 1.

Ad libitum water was supplied and daily 130 gm of mash feed was offered to the layer hens throughout the experimental period. 14 hours and 10 hours dark period was provided. KMNO4 solution was used in foot bath for biosecurity.

Data recording and sample collection

Number of eggs laid by each treatment was recorded daily and the eggs were weighed just after collection. Three eggs from each replication was collected weekly during the whole period of the experiment and used as sample for analysis.

Table 1. Chemical composition of Diet

Feed ingredients

Amount (%)



Protein concentrate


Rice polish


Soya meal






Soyabean oil


Vit-min premix








Calculated composition g/100gm Dry matter

Crude protein


Crude fiber


Total Phosphorus










ME (Kcal/kg)


DCP = Di–calcium phosphate; Vit-Min Premix = Vitamin-Mineral premix; A. Phosphorus= Available Phosphorus; ME= Metabolizable energy; Vitamin-mineral premix composition: Vitamin A 12,500,000 IU, Vitamin D3 2,500,000 IU, Vitamin E 20,000 mg, Vitamin K3 4,000 mg, Iron 40,000 mg Vitamin B1 2,500 mg, Vitamin B2 5,000 mg, Vitamin B6 4,000 mg Nicotinic acid 40,000 mcg, Pantothenic acid 12,500 mg, Vitamin B12 12,000 mcg, Folic acid 800 mg, Biotin 100 mg, Cobalt 400 mg, Copper 10,000 mg, Iodine 400 mg, Manganese 60,000 mg, Zinc 50,000 mg, Selenium 150 mg, Di-Calcium Phosphate 380 gm

Calculation of production performance and Egg quality

Egg weight was measured using scale of 0.1 gm accuracy. Hen day egg production and Egg mass output was measured using the following formula:

Hen day egg production= (Number of eggs laid on a day/number of birds housed on a day) ×100

Egg mass output (g/egg/hen/day) = (Egg weight × Hen day egg production)/100

Haugh unit (HU) score was calculated according to Haugh (1973),

HU= 100 log (h-1.7W0.37 + 7.6)

Where, h= average thick albumen height (mm), W= egg weight (g)

Other egg quality was determined using the following formula:

A caliper was used to measure the egg height, albumen and yolk heights as well as egg and egg yolk diameter.

Statistical analysis

To examine the significance of treatment effects, all recorded and calculated data were subjected to analysis of variance (ANOVA) in a Completely Randomized Design (CRD). Tukey's HSD test was used to compare the treatment means. SPSS statistical analysis software was used for all of these statistical analyses (SPSS Inc. Chicago, IL, USA). The comparison of means was done at a significance level of 5%.


Production trait of laying hen

There was significant relationship of supplementing spirulina on production performance of laying hens. Hen day egg production was numerically high (12.19%) in 0.2% than control group (0%) (Figure 1). Significantly maximum weight of egg was observed in the dietary treatment group of 0.2% and lowest was in the group of 0% (Table 2).

Table 2. Production performance of laying hen

of Spirulina

Hen Day Egg

weight (g)

Egg mass output
(g/ egg/hen/day)

















p value




0%= Control, 0.1%=Control+0.1% Spirulina, 0.2%=Control+0.2% Spirulina, 0.3%=Control+0.3% Spirulina;
abcmeans bearing dissimilar superscript in a column differ significantly at the level of 5%

Egg mass output was significantly maximum (14.53% than control group) in 0.2%. More than 60 g/ egg/hen/day egg mass output was found only in the dietary treatment group of 0.2% and 0.3%.

Egg quality

Among the dietary treatment group external quality of egg was not significant different except shell thickness. Shape index was numerically high in 0.1% and 0.2% than 0% and 0.3%. Shape index was 3.63% higher in 0.2% than control group (Table 3).

Table 3. External quality of egg

of Spirulina

Index (%)

Thickness (mm)

% Shell

















p value




0%= Control, 0.1%=Control+0.1% Spirulina, 0.2%=Control+0.2% Spirulina, 0.3%=Control+0.3% Spirulina;
means bearing dissimilar superscript in a column differ significantly at the level of 5%

Shell thickness 8.3 mm and 8.4 mm was in the dietary treatment group of 0.1% and 0.2% respectively which was more or less 18.30% high than control group. % Shell was also high in 0.2% (0.42) than control group. Maximum (p<0.05) albumen index was found in 0.2% (8.60% than control group 5.30%) (Table 4).

Figure 1. Hen day egg production at different level of spirulina supplementation

Table 4. Internal Quality of egg

of Spirulina

Albumen Index (%)

Haugh Unit

Yolk Index (%)

















p value




0%= Control, 0.1%=Control+0.1% Spirulina, 0.2%=Control+0.2% Spirulina, 0.3%=Control+0.3% Spirulina; abc means bearing dissimilar superscript in a column differ significantly at the level of 5%

Haugh unit was 82.20 which was 12.0% high (p<0.05) in 0.2% group than 0%. In case of yolk color, L* was maximum (35.01) in 0% and lowest (30.82 and 30.22) in 0.2% and 0.3% respectively (Table 5).

Table 5. LAB

of Spirulina




















p value




0%= Control, 0.1%=Control+0.1% Spirulina, 0.2%=Control+0.2% Spirulina, 0.3%=Control+0.3% Spirulina; abc means bearing dissimilar superscript in a column differ significantly at the level of 5%
a* and b* was also maximum in 0.2% than control group and in the group of 0.3% have a* and b* value which was close to 0.2%.


Egg production was not significantly affected by the supplementation of spirulina which was supported by the findings of Zahroojian (2013) who used different level of spirulina in layer diet. Weight of egg was high in spirulina supplemented group and this results are in the agreement with Mariey (2012) who reported that inclusion of spirulina in layer diet increase egg weight and egg mass output. This increase of egg weight could be due to high protein content of spirulina (Omri et al 2019). Park (2015) showed that supplementation of marine microalgae 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). Dogan et al (2016) reported that, percent egg shell increased with increasing supplementation of Spirulina platensis from 5 to 20 g/kg diet. This increase may be due to the high calcium content in Spirulina platensis (Tokusoglu and Unal, 2003) which supports the present experiment. Parisse (2014) stated that egg having Haugh unit more than 70 are of excellent quality and in the present experiment eggs of 0.2% spirulina supplemented group had Haugh unit 82.20 which indicates higher quality of egg. Supplementation of 0.15% spirulina increase egg yolk weight (Mariey et al 2012). Laying hens are unable to produce pigment in egg yolk and need to provide these through their diet (Park, 2015). Phytopigments like phycobilins, phycocyanin and allophycocyanin are present in Spirulina platensis (Bermejo 2008; Shimkus 2009; Hazati 2019). So, dietary inclusion of spirulina could be one of the best ways to provide phytopigments which enhance the yolk coloration and fulfill the consumer’s preference. Supplementation of spirulina increases the egg yolk color (Zahroojian, 2013; Selim, 2018). Spirulina’s effect on yolk colour results from its high level content of zeaxanthin, xanthophylls and other carotenoid pigments, particularly β-carotene which accumulate within the yolk (Anderson, 1991; Takashi, 2003; Kotrbacek, 2013). It's likely that Spirulina's carotenoids are well absorbed by the bird's gastrointestinal tract and accumulate in a variety of places, including the egg yolk, where they're mostly deposited as free carotenoids (Latscha, 1990). Thus, spirulina supplementation through diet may enhance egg external and internal quality as well as egg yolk pigmentation.



Abdel-Daim M M, Farouk S M, Madkour F F and Azab S S 2015 Anti-inflammatory and immunomodulatory effects of Spirulina platensis in comparison to Dunaliella salina in acetic acid-induced rat experimental colitis. Immunopharmacology Immunotoxicology, 37:126-139.

Anderson D W, Tang C and Ross E 1991 The xanthophylls of Spirulina and their effect on egg yolk pigmentation. Poultry Science, 70: 115-119.

Bermejo P, Pinero E and Villar A M 2008 Iron-chelating ability and antioxidant properties of phycocyanin isolated from a protean extract of Spirulina platensis. Food Chemistry, 110: 436- 445.

Dismukes G C, Carrieri D, Bennette N, Ananyev G M and Posewitz M C 2008 Aquatic phototrophs: efficient alternatives to land-based crops for biofuels. Current Opinion in Biotechnology, 19: 235– 240.

Dogan S C, Baylan M, Erdogan Z, Akpinar G C, Kucukgul A and Duzguner V 2016 Performance, egg quality and serum parameters of Japanese quails fed diet supplemented with Spirulina platensis. 25(12): 5857-5862.

Doreau M, Bauchart D and Chilliard Y 2010 Enhancing fatty acid composition of milk and meat through animal feeding. Animal Production Science, 51: 19– 29.

Güroy B, Şahin I and Mantoğlu S 2012 Spirulina as a natural carotenoid source on growth, pigmentation and reproductive performance of yellow tail cichlid Pseudotropheus acei. Aquaculture International, 20: 869–878.

Hajati H and Zaghari M 2019 Effects of Spirulina platensis on Growth Performance, Carcass Characteristics, Egg Traits and Immunity Response of Japanese Quails. Iranian Journal of Applied Animal Science, 9(2): 347-357

Haugh R R 1973 The Haugh unit for measuring egg quality. U. S. Egg Poultry Magazine, 43:552–555.

Jamil A B M, Akanda M, Rahman M, Hossain M and Islam M 2015 Prebiotic competence of spirulina on the production performance of broiler chickens. Journal of Advanced Veterinary and Animal Research, 2(3): 304-309.

Kotrbacek V, Skrivan M, Kopecky J, Penkava O, Hudeckova P, Uhrikova I and Doubek J 2013 Retention of carotenoids in egg yolks of laying hens supplemented with heterotrophic Chlorella. Czech Journal of Animal Science, 58: 193-200.

Mariey Y, Samak A H R and Ibrahem M A 2012 Effect of using spirulina platensis algae as a feed additive for poultry diets: Productive and reproductive performances of local laying hens. Egyptian Poultry Science, 32:201–215.

Omri B, Amraoui M, Tarek A, Lucarini M, Durazzo A, Cicero N, Santini A and Kamoun M 2019 Arthrospira Platensis (Spirulina) Supplementation on Laying Hens’ Performance: Eggs Physical, Chemical, and Sensorial Qualities. Foods, 8: 1-12.

Parisse A 2014 Évolution qualitative et quantitative des composantes de l’oeuf pendant les trois phases de ponte chez la poule. Journal of Applied Bioscience, 74: 6080–6085. DOI: 10.4314/jab.v74i1.9

Park J H, Upadhaya S D and Kim I H 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-Austra. Journal of Animal Science, 28: 391–397.

Park J H, Upadhaya S D and Kim I H 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. Journal of Animal Science 28: 391-397. DOI: 10.5713/ajas.14.0463

Selim S, Hussein E and Abou Elkhar R 2018 Effect of Spirulina platensis as a feed additive on laying performance, egg quality and hepatoprotective activity of laying hens. European Poultry Science, 82: 1–14

Shimkus A, Shimkiene A, Juozaitiene V, Zavodnik L, Juozaitis A and Muzikevicius A 2009 Influence of blue algae spirulina platensis on the productivity of sows. Dokladi na B lgarskata Akademiâ na Naukite. 62: 405-410.

Sujatha T and Narahari D 2011 Effect of designer diets on egg yolk composition of ‘White Leghorn’ hens. Journal of Food Science and Technology, 48: 494–497.

Takashi S 2003 Effect of administration of Spirulina on egg quality and egg components. Animal Husbandry, 57: 191– 195.

Tokusoglu O and Unal M K, 2003 Biomass nutrient profiles of three microalgae: Spirulina platensis, Chlorella vulgaris, and Isochrisis galbana. Journal of Food Science, 68: 1144–1148.

Zahroojian N, Morave H and Shivazad M 2013 Effects of dietary marine algae (Spirulina platensis) on egg quality and production performance of laying hens. Journal of Agricultural Science and Technology,15: 1353–1360.