You are on page 1of 6

The Effects of Different Levels Chlorell

Microalgae on Performance and
Immune Response of Laying Hens
under Heat Stress Condition
Nasroallah Moradikor, Neda Mohamadi

International Journal of Life Sciences
ISSN No. 2091-0525

OPEN

ACCESS

DOIdx.doi.org/10.3126/ijls.v9i2.12058

This article is protected by copyright, and all rights are held exclusively by International Journal of Life Sciences. © IJLS

International Journal of Life Sciences 9 (2) : 2015; 71 - 74
Founded 2007

An Independent, Open Access, Peer Reviewed, Non-Profit Journal

International Journal of Life Sciences
ISSN 2091-0525

website: http://nepjol.info/index.php/IJLS/index

International Journal of

Life Sciences
Copyright © International Journal of Life Sciences

Research Article

The Effects of Different Levels Chlorell Microalgae on
Performance and Immune Response of Laying Hens under Heat
Stress Condition
Nasroallah Moradikor*, Neda Mohamadi
Young Researchers and Elites Club, Kerman Branch, Islamic Azad University, Kerman, Iran

Article Information

ABSTRACT

Key words:
Microalgae Chlorell;
Performance;
Immune response;
Laying hens

This study was conducted to determine The Effects of different levels microalgae Chlorell sp. on
performance of laying hens under heat stress condition. A completely randomized design
experiment, with 7 replicates and 6 treatments (9 birds per replicate) was carried out by
allotting 378 white Lohman strain laying hens at 65 weeks of age. Six experimental trearments
were continual addition of 0, 100, 200, 300, 400 ppm microalgae Chlorell and 400 ppm toein to
drinking water were fed to laying hens for 9 weeks. Egg production, egg weight, egg mass output
and feed intake were determined. Antibody production against sheep red blood cells (SRBC)
also was measured. Data were analyzed by ANOVA using General Linear Models procedure
of SAS software and comparison of means were compared using Duncan's multiple range
test. The results of this experiment showed that microalgae Chlorell had no effect on productive
performance. Adding microalgae Chlorell to the drinking water improved immune response of
laying hens and a more positive effect was observed when 400 ppm added to the water. It is
concluded that the addition of microalgae Chlorell to the water drinking of laying hens
improved immune response of laying hens.

balanced nutrition and improve animal growth, several
reports advise mixing species from the ones listed
The use of microalgae as a feed additive could be the best above. In poultry rations, algae up to a level of 5-10% can
choice for solving problems associated with use of be used safely as partial replacement for conventional
antibiotics, organic acids and other chemical ingredients proteins (Spolaore et al, 2006). The yellow colour of
in feed. Microalgae are required for larval nutrition broiler skin and shanks as well as of egg yolk is the most
during a brief period, either for direct consumption in the important characteristics that can be influenced by
case of molluscs and peneid shrimp or indirectly as food feeding algae (Becker, 2004). Moreover, the Institut fü r
for the live prey fed to small fish larvae (Brown et al., Getreideverarbeitung (Bergholz-Rehbrü cke, Germany)
1997, Muller-Feuga, 2000). In addition to its use in produces a natural feed with the algae Chlorella and
human nutrition, microalgae can be incorporated into Arthrospira called Algrow. Ginzberg et al. (2000) studied
the feed for a wide variety of animals ranging from fish role of algae, Porphyridium sp. as feed supplement on
(aquaculture) to pets and farm animals. In fact, 30% metabolism of chicken. Earlier results in the same
of the current world algal production is sold for animal laboratory showed a reduction in serum cholesterol and
feed applications (Becker, 2004) and over 50% of the triglyceride levels in rodents fed with red algal biomass.
current world production of Arthrospira is used as feed In this study lyophilized algae biomass was fed to
supplement (Yamaguchi, 1997). To provide more better chickens at a proportion of 5% or 10% of the standard

INTRODUCTION

*Corresponding author Email address: moradikor.nasroallah@yahoo.com
Cite this article as Moradikor, N.; Mohamadi, N. (2015) The Effects of Different Levels Chlorell Microalgae on Performance and Immune Response of
Laying Hens under Heat Stress Condition. International Journal of Life Sciences 9(2): 71 - 74.
Doi:dx.doi.org/10.3126/ijls.v9i2.12058
Copyright reserved © International Journal of Life Sciences

DOI:dx.doi.org/10.3126/ijls.v9i2.12058

International Journal of Life Sciences 9 (2) : 2015; 71 - 74

chicken diet. Chickens fed with algae biomass consumed
10% less food and their serum cholesterol levels were
significantly lower (by 11% and 28% for the groups fed
with 5% and 10% supplement, respectively) as
compared with the respective values of the control group
(with unsupplemented diet). Egg yolk of chickens fed
with algae tended to have reduced cholesterol levels (by
10%) and increased linoleic acid and arachidonic acid
levels (by 29% and 24%, respectively). In addition the
colour of egg yolk became darker, indicating that higher
carotenoid was produced (2.4 fold higher). Other poultry
feeding studies with Spirulina (up to 30%) showed that
both protein and energy efficiency of this alga were
similar to other conventional protein carriers up to a level
of 10% (Becker, 2004). Significantly higher growth rates
and lower non-specific mortality rate were observed in
turkey poults fed with Spirulina at the level of 1-10 g.kg -1
diet. Waldenstedt et al. (2003) analyzed the effects of a
dietary astaxanthin from the microalgae Haematococcus
pluvialis on broiler chickens performance. The results
indicated that tissue astanxanthin and carotenoid
concentrations increased with increasing levels of algal
meal inclusion. The results also indicate that algal meal
could reduce caecal colonization of Clostridium
perfringes. This gives better results than a diet
composed of only one algal species (Yamaguchi, 1997).
While microalgae provide food for zooplanktons, they
also help to stabilize and improve the quality of the
culture medium. Indeed, for numerous freshwater and
seawater animal species, the introduction of
phytoplanktons to rearing ponds (green-water
technique) leads to much better results in terms of
survival, growth and transformation index than that of
the clear-water technique (Lio-Po et al., 2005). The
reasons for this are not entirely known but may include
(Muller-Feuga, 2004) water quality improvement and
stabilization by algal oxygen production and pH
stabilization, the action of some excreted biochemical
compounds along with the induction of behavioral
processes like initial prey catching, and the regulation of
bacterial population, probiotic effects (Irianto and
Austin, 2002), and the stimulation of immunity
(suggested but not sufficiently understood). Many
nutritional and toxicological evaluations have proved the
suitability of algal biomass as feed supplement (Becker,
2004). Arthrospira is largely used in this domain and
concerns many types of animal: cats, dogs, aquarium
fish, ornamental birds, horses, cows and breeding
bulls. Algae positively affect the physiology (by
providing a large profile of natural vitamins, minerals,
and essential fatty acids; improved immune response
and fertility; and better weight control) and their
external appearance (resulting in healthy skin and a
lustrous coat) of animals. In poultry rations, algae up to
a level of 5–10% can be used safely as partial replacement
for conventional proteins.The present study was
conducted to examine the effects of different levels
Chlorell microalgae on performance and Egg quality
traits of laying hens under heat stress condition.

MATERIALS AND METHODS
A completely randomized design experiment, with 7
replicates and 6 treatments (9 birds per replicate) was
carried out by allotting 378 white Lohman strain laying
hens at 65 weeks of age. Six experimental trearments
were continual addition of 0, 100, 200, 300, 400 ppm
microalgae Chlorell and 400 ppm toein to drinking water
were fed to laying hens for 9 weeks. Before start of the
experiment, all hens fed basal diet for 2 weeks and were
similar in body size and production. During the
experiment, hens fed approximately 120 (gr/day) and
water was offered ad-libitum. The hen's performance
including hen-day egg production %, feed intake and egg
mass (g/hen/day) was measured. Feed conversion (FCR,
g feed: g egg) was also calculated as the ratio of gram of
feed consumed per gram of egg weight produced. At the
6th week of the experiment, 6 hens were randomly
selected from each group (1 from each replicate) and
injected with 0.2 ml of 9% suspension of sheep
erythrocytes (SRBC) in phosphate buffer saline. One
week after SRBC injection, 3 mL blood was taken from
selected hens using jugular venipuncture, and serum was
s e p a r a t e d a n d e v a l u a t e d f o r a n t i b o d y t i t e r.
Haemagglutination inhibition (HI) test was used for
determining antibody titer sera.

Statistical Analysis:
Data were analyzed by ANOVA using General Linear
Models procedure of SAS software. The differences
among treatment means determined using the Duncan's
multiple-range test. A probability value of P ≤ 0.05
indicated that the difference was statistically significant.
Linear and quadratic polynomial contrasts were used to
evaluate the effect of different levels of Chlorell
microalgae.

RESULTS AND DISCUSSION
The effect of different levels of supplemental Chlorell
microalgae to drinking water on egg production and egg
mass of laying hens during the entire 9-week period of
the study are presented in Table 1. The results indicated
that inclusion of Chlorell microalgae had no significant
effect on egg production and egg mass (P>0.05). Egg
production and egg mass have not changed with
increasing the levels of Chlorell microalgae.
The effects of different levels of supplemental Chlorell
microalgae on feed intake and feed conversion ratio of
laying hens during the entire 9-week period of the study
is shown in Table 2. The results indicated that inclusion of
Chlorell microalgae have not a significant effect on feed
conversion ratio (P>0.05). In addition, inclusion of
Chlorell microalgae had not significant effect on feed
intake (P>0.05). Feed intake was lower in T5 treatment

72

International Journal of Life Sciences 9 (2) : 2015; 71 - 74

compared with other treatments. In addition, water
supplementation with Chlorell microalgae decreased
feed conversion ratio.
Table 1: The effects of different levels Chlorell microalgae
supplementation on egg production and egg mass of laying
hens

Table 2: The effects of different levels Chlorell microalgae
supplementation on feed intake and Feed conversion ratio
of laying hens

The effects of different levels of supplemental Chlorell
microalgae to drinking water on egg weight and antibody
titer against SRBC of laying hens during the entire 9-week
period of the study is shown in Table 3. Results from this
table indicated that inclusion of Chlorell microalgae had
a significant effect on antibody titer against SRBC
(P<0.05). Egg weight increased with increasing Chlorell
microalgae to drinking water. The hens receiving the 400
ppm microalgae Chlorell to drinking water had
significantly higher titers of total, IgM and IgG antibodie
than that of other treatments (P<0.05). The titer of total
antibody, IgM and IgG significantly increased with
increasing Chlorell microalgae to drinking water.

Table 3: The effects of different levels Chlorell microalgae
supplementation on egg weight and antibody titer against
SRBC of laying hens

Results from current study indicated that inclusion of
microalgae Chlorell to drinking water had not significant
effect on production performance of laying hens
(P>0.05). But Chlorell microalgae to the drinking water
improved immune response of laying hens. It is an
attractive potential food source because it is high in
protein and other essential nutrients; when dried, it is
about 45% protein, 20% fat, 20% carbohydrate, 5% fibre,
and 10% minerals and vitamins. Mass-production
methods are now being used to cultivate it in large
artificial circular ponds. It is also abundant in calories, fat,
and vitamins (Warren, 1997). When first harvested,
Chlorella was suggested as an inexpensive protein
supplement to the human diet. Advocates sometimes
focus on other supposed health benefits of the algae, such
as claims of weight control, cancer prevention, and
immune system support (Warren, 1997). One small (35
participant) study suggested Chlorella supplementation
has a positive effect on the reduction of dioxin levels in
breast milk and it may also have beneficial effects on
nursing infants by increasing the IgA levels in breast milk
(Nakano et al., 2007). The most important substance in
C h l o re l l a i s β - 1 , 3 - g l u c a n , wh i c h i s a n a c t ive
immunostimulator, a free radical scavenger and a
reducer of blood lipids (9; Ryll et al., Abstr. Eur. Workshop
Microalgal Biotechnol., Germany, p. 56, 2003). However,
various other health-promoting effects have been
clarified (efficacy on gastric ulcers, wounds, and
constipation; preventive action against atherosclerosis
and hypercholesterolemia; and antitumor action) (JongYuh and Mei-Fen, 2005). Chlorella can also be used as a
food additive owing to the taste and flavour-adjusting
actions of its coloring agent (Gouveia et al., 1996). The
study of different aspects related to the behavior of
microalgae has received renewed interest due to the wide
field of application of these microorganisms. Algae
cultures have been principally developed as an important
source of many products, such as aquaculture feeds,
human food supplements, and pharmaceuticals and they
have been suggested as a very good candidate for fuel
production (Pulz and Gross, 2001). In the end, scientists
discovered that Chlorella would be much more difficult to
produce than previously thought. The experimental
research was carried out in laboratories, not in the field.
In order to be practical, the entire batch of algae grown
would have to be placed either in artificial light or in
shade to produce at its maximum photosynthetic
efficiency. Also, for the Chlorella to be as productive as the
world would require, it would have to be grown in
carbonated water, which would have added millions to
the production cost. A sophisticated process, and
additional cost, was required to harvest the crop, and, for
Chlorella to be a viable food source, its cell walls would
have to be pulverized. The plant could reach its
nutritional potential only in highly modified artificial
situations. Another problem was developing sufficiently
palatable food products from Chlorella (Becker, 2007).
Although the production of Chlorella looked promising
and involved creative technology, it has not to date been

73

International Journal of Life Sciences 9 (2) : 2015; 71 - 74

cultivated on the scale some had predicted. It has not
been sold on the scale of Spirulina, soybean products, or
whole-grains. Costs have remained high, and Chlorella
has for the most part been sold as a health food, for
cosmetics, or as animal feed (Becker, 2007).

REFERENCES
Apt, K. E. and Behrens, P. W. (1999). Commercial developments in microalgal
biotechnology. J. Phycol., 35, 215-226.
Becker EW. 2004. Microalgae in human and animal nutrition. In: Richmond A., editor.
Handbook of Microalgae Culture. Biotechnology and Applied Phycology. Oxford:
Blackwell Science.
Becker, E.W. (2007). Micro-algae as a source of protein. Biotechnology Advances 25 (2):
207-210.

CONCLUSION
The present investigation suggested that supplemental of
Chlorell microalgae to drinking water at a level of 400
ppm improve immune responses of laying hens. Thus,
supplementation of Chlorell microalgae at levels above
recommended as nutritional requirements for improve
humoral and cellular immunity. In addition, improving
immune responses of laying hens by Chlorell microalgae
supplementation is relatively a novelty results, so the
more effect of Chlorell microalgae could be the subject of
further investigations.

Belasco, Warren (July 1997). Algae Burgers for a Hungry World? The Rise and Fall of
Chlorella Cuisine. Technology and Culture, 38 (3): 608-34.
Brown, M. R., Jeffrey, S. W., Volkman, J. K., and Dunstan,G. A. (1997).Nutritional
properties of microalgae for mariculture. Aquaculture, 151, 315-331.
Chuntapa, D., Powtongsook, S., and Menasveta, P. (2003). Water quality control using
Spirulina platensis in shrimp culture tanks. Aquaculture, 220, 355-366.
Ginzberg A. Cohen M. Sod-Mariah U.A. Shany S. Rosenshtrauch A. e Arad S.2000.
Chickens fed with biomass of the red microalga Porphyridium sp. have reduced blood
cholesterol level and modified fatty acid composition in egg yolk. Journal of applied
phycology 12: 325-330.
Gouveia, L., Veloso, V., Reis, A., Fernandes, H., Novais, J., and Empis, J., (1996). Evolution
of pigment composition in Chlorella vulgaris. Bioresour. Technol., 57, 157-159.
Irianto, A. and Austin, B. (2002). Probiotics in aquaculture. J. Fish Dis., 25, 633–642.
Jong-Yuh, C. and Mei-Fen,S., (2005). Potential hypoglycemic effects of Chlorella in
streptozotocin induced diabetic mice. Life Sci., 77, 980-990.
Lio-Po, G. D., Leañ o, E. M., Peñ aranda, M. M. D., Villa-Franco, A. U., Sombito, C. D., and
Guanzon, N. G. (2005). Antiluminous Vibrio factors associated with the 'green water'
grow out culture of the tiger shrimp Penaeus monodon. Aquaculture, 250, 1-7.

ACKNOWLEDGMENTS
This research was funded and supported by research
budget (Research Project Code= 91580) of Young
Researchers and Elite Club, Kerman Branch, Islamic Azad
University, Kerman, Iran. Their contribution is gratefully
acknowledged. Also we would like to have a special thank
to Dr. Peyman Rajaei for providing a friendly
environment, kind helps and research facilities.

Muller-Feuga, A.: Microalgae for aquaculture, (2004). The current global situation
and future trends, p. 352–364. In Richmond, A. (ed.), Handbook of microalgal culture.
Blackwell, Oxford.
Muller-Feuga, A. (2000). The role of microalgae in aquaculture: situation and trends. J.
Appl. Phycol., 12, 527-534.
Nakano, Shiro; Takekoshi, Hideo; Nakano, Masuo (2007). Chlorella(Chlorella
pyrenoidosa) Supplementation Decreases Dioxin and Increases Immunoglobulin a
Concentrations in Breast Milk. Journal of Medicinal Food 10 (1): 134-42.
PULZ O., GRASS W. (2001).Valuable products from biotechnology of microalgae. Applied
Microbiology and Biotechnology, Vol. 65, pp. 635-648.
Rodolfi, L., Chini Zittelli, G., Barsanti, L., Rosati, G., and Tredici, M. R. (2003). Growth
medium recycling in Nannochloropsis sp. mass cultivation. Biomol. Eng., 20, 243-248.
Spolaore P., Joannis-Cassan C., Duran E. and Isambert A. 2006. Commercial applications
of microalgae. Journal of Biosciences and Bioengineering 101 (2): 86-96.

© International Journal of Life Sciences
NOTE: Authors/contributors are responsible
for originality, contents, correct references, and
ethical issues.

Waldenstedt, L., Inborr, J. Hansson, I. Elwinger, K. 2003. Effects of astaxanthin-rich algal
meal (Haematococcus pluvalis) on growth performance, caecal campylobacter and
clostridial counts and tissue astaxanthin concentration of broiler chickens. Animal Feed
Science and Technology 108: 119-132.
Yamaguchi, K. (1997). Recent advances in microalgal bioscience in Japan, with special
reference to utilization of biomass and metabolites: a review. J. Appl. Phycol., 8, 487502.

Submit your next manuscript to IJLS with a 1. Convenient online submission,
2. Rapid editorial review followed by peer review,
3. Immediate publication on acceptance.
74