Scientific Papers. Series D. Animal Science. Vol.

LXI, Number 2, 2018

ISSN 2285-5750; ISSN CD-ROM 2285-5769; ISSN Online 2393-2260; ISSN-L 2285-5750

SOURCES OF CAROTENOIDS AND THEIR USES

AS ANIMAL FEED ADDITIVES – A REVIEW

Diana PASARIN, Camelia ROVINARU

National Institute for Research and Development in Chemistry and Petrochemistry

202 Spl. Independentei, Bucharest, Romania

Corresponding author email: rovinarucamelia@yahoo.com

Abstract

Carotenoids are natural pigments, widely distributed in nature, synthesized by plants, algae, fungi, and phototrophic
bacteria. Carotenoids have coloring power and antioxidant properties, being used as colorants for foods, cosmetics and
feeds. Nowadays, a small part of industrial carotenoids is obtained from algae and plants, and the most part is
chemically synthesized, but with high production cost. Taking into account consumer’s demand for natural compounds,
there is an increased trend for products containing natural ingredients including those used in animal feed. Products
quality and animal health are influenced by carotenoids added in feed as important component in daily diet. This paper
reviews aspects regarding the main sources of carotenoids used as feed additives for nutritional and coloring
properties and their impact on animal and human health.

Key words: carotenoids, feed additives, sources of carotenoids, pigmentation.

INTRODUCTION properties and deposition of carotenoids crucial

for consumers (egg yolks and other products).
Carotenoids gained great interest lately for food In fact, as studies show, there is a strong
and feeding, as their functions related to relationship between diets/foods rich in
nutrition, health and wellbeing are well carotenoids and the risk reduction of various
appreciated. As consumers become more and diseases. Carotenoids increase the immune
more aware of the importance of natural response, are excellent antioxidants, scavenging
instead of synthetic and artificial ingredients, and inactivating free radicals. This is why both
they tend to change preferences and replace vitamin A precursor carotenoids and the non-
products in their diets according to norms precursors ones play an important role in the
which no longer respond to good, tasty and protection and inhibition of serious diseases as
affordable consumptions, yet to higher, long- cancer, atherosclerosis, cataracts, macular
term perspective standards (García-Chavarría degeneration, multiple sclerosis, degenerative
and Lara-Flores, 2013; Carotenoids Market by diseases, and cardiovascular diseases
Type, 2016; Mohiseni., 2017). (Mezzomo et al., 2016).
In this line, arguments related to the On the other hand, there is no novelty feeding
replacement of synthetic carotenoids with animals with carotenoids, these have always
natural ones in case of animal feed additives been part of their daily diet. Astaxanthin is the
lay in multiple health impact through major carotenoid used for pigmentation of fish
environment and animal to human health (Kaur muscle, mainly salmons (Bhosale, 2004). Beta-
and Shah, 2017). carotene, lycopene, and lutein increase growth
As such, many researches have been directed to performance of animals and quality of
in-depth knowledge on carotenoid sources, pigmented products (Nelis and De Leenheer,
especially natural ones, for their use as animal 1991). Animals and humans are unable to
feed additives, so as to benefit from their synthesize carotenoids, they have to meet their
functions including skin and tissues daily nutritional needs from the products
pigmentation (especially when it comes to ingested (Priyadarshani and Rath, 2012).
animals/birds grown in captivity), as growth Carotenoids, as part of the nutrients in the feed,
promoters (in terms of mass production), for support animal health and animal products
reproduction and survival goals, sensory quality (Amaya et al., 2014). They have an
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important role in molecular processes of cell molecules, joined in a head to tail pattern
membranes whose structure, properties and (Mattea, 2009; Domonkos, 2013). Structurally,
stability can be modified, leading to possible carotenoids take the form of a polyene chain
beneficial effects on human health (Zaheer, that functions as a chromophore, due to 9-11
2017). conjugated double bonds and possibly
Out of high production and marketability terminating in rings, what determines their
reasons, carotenoids are present in the animal characteristic color in the yellow to red range
kingdom, playing functions such as coloring (Vershinin, 1999). The presence of different
(pets/ornamental birds and fish, mimicking), number of conjugated double bounds leades to
flavoring (scents and pollination in nature), various stereoisomers abbreviated as E- and Z-
reproduction (bird feathers and finding mates; isomers, depending on whether the double
development of embryos), resistance to bonds are in the trans (E) or cis (Z)
bacterial and fungal diseases, immune configuration (Vincente et al., 2017).
responses (lutein connected to anti- Carotenoids are synthesized by all
inflammatory natural substance in poultry), as photosynthetic organisms and some non-
well as normal development of skin and photosynthetic bacteria and fungi (Ruiz-Sola,
mucosa. 2012). Due to length of the polyene
The aim of this review is to highlight the tail, carotenoid compounds are involved in
various sources of carotenoids and their capturing energy from light, absorbing
application in animal feed additives. This topic wavelengths, ranging from 400-550
has rarely been described in a general matter, nanometers (violet to green light), thus
and as such we endeavor to provide an assisting in photosynthesis and causes the
overview of published works on the subject. compounds to be deeply colored yellow,
orange, or red (Vershinin, 1999; Zakynthinos,
DEFINITION, CHEMICAL STRUCTURE 2016).
AND CLASSIFICATION OF There have been identified over 750 naturally
CAROTENOIDS occurring carotenoids, biosynthesized de novo
by plants, algae, fungi and bacteria (Okada et
Carotenoids are natural pigments, widely al., 2008), among which only 50 have
distributed in nature, responsible for the significant biological activity for animals and
coloring of fruits, vegetables and flowers. humans (Mezzomo et al., 2016).
Carotenoids take their name from carrot Animals cannot synthesize carotenoids (Rock,
(Daucus carota), which accumulates high 2009), obtaining them through dietary intake,
levels of these pigments in the root, being an with a few exceptions, represented by aphids
exception among plants (Stange, 2015). They and spiders which acquired the ability and
are used as food colorants, feed supplements, genes from fungi (Moran, 2010; Nováková,
for cosmetic and pharmaceutical purposes. 2012) or are produced by endosymbiotic
Most carotenoids are made up of eight bacteria in whiteflies (Sloan, 2012).
condensed 5-carbon-containing isoprene Carotenoids are classified in Table 1.

Table 1. Classification of carotenoids

Classification
Major classes Structure Examples Attribute References
criteria

Chemical Carotenes Hydrocarbons (constituted α-carotene -color range from pale Delgado-Vargas, 2000
structure by carbon and hydrogen) β- carotene yellow, bright orange to Tinoi, 2005
that can be cyclized at one γ-carotene deep red Aizawa, 2007
or both ends of the molecule lycopene -maintains eye health Kaur and Shah, 2017
torulene -does not color the fish

Oxycarotenoids Oxygenated hydrocarbon lutein, zeaxanthin, β- -generally yellow in color Delgado-Vargas,

(xanthophylls) derivatives that contain at cryptoxanthin, -antiallergic and anti- 2000;
least one oxygen atom as astaxanthin, cancerous actions Bhosale and Berstein,
hydroxyl groups, keto, canthaxanthin, -can color the fish 2005;
epoxy, methoxy, or violaxanthin,neoxanthin, -colors egg yolks Kushwaha et al, 2014.
carboxylic acid antheraxanthin

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 Classification
Major classes Structure Examples Attribute References
criteria

Functionality Primary Xanthophylls β-carotene -structural and functional Delgado-Vargas, 2000

violaxanthin neoxanthin components of the cellular
photosynthetic apparatus

Secondary α-carotene -localized in fruits and Delgado-Vargas, 2000

β-cryptoxanthin flowers
zeaxanthin -encompasses carotenoids
antheraxanthin capsanthin produced at large levels
capsorubin

Nutritionally Precursors of Have at least one ring of β- α-carotene -helps maintain eye Ambrosio et al., 2006;
vitamin A ionone not replaced and side β-carotene health, healthy mucus Premkumar, 2014
polyenic chain with at least β-cryptoxanthin membranes and immunity
11 carbons

Non lutein Premkumar, 2014

provitamin A zeaxanthin
lycopene

β-carotene is used as food colorant and food ruberrimus. Its structural characteristics make
supplement having antioxidant capacity and possible antitumoral and immune enhancing
provitamin A activity, in a concentration ranges activities and could be used as food, feed and
from 2 to 50 ppm. Once converted to vitamin cosmetic additives (Zoz et al, 2015).
A, health benefits include maintenance of Lutein is a dihydroxy-carotene with an alcohol
normal eye health, epithelial function, group containing hydroaromatic α structure
embryonic development, and immune system (Kim, 2006) being in covalent interactions
function. Currently, the application of within fatty acids (Mezzomo, 2016). Lutein is
carotenoids in food formulations is limited found in leaves of green vegetables (spinach,
because of it poor water solubility, high kale, pumpkin, turnip greens) and is yellow-
melting point and chemical instability (Qian, colored. Lutein gives color to chicken fat, egg
2012a; Piorkowski, 2014), promoted by heat, yolk, and chicken feathers and also increase the
light, singlet oxygen, acid, iron and iodine, and efficacy of vaccination against infectious
free radical, because of conjugated polyene bronchitis virus in laying hens (McGraw,
chain which is characteristic of carotenoids 2003).
(Dutta, 2005; Boon, 2010). Good sources of Lutein and zeaxanthin are associated with eye
beta-carotene are cantaloupe, mangoes, papaya, health, because they are the only carotenoids
carrots, sweet potatoes, spinach, kale and found in the retina in macula lutea, being likely
pumpkin (Premkumar, 2014). effective in age-related macular degeneration
Lycopene is an aliphatic carotenoid which (Premkumar, 2014).
dissolves in the oil, and that is why the Astaxanthin is a dark-red organic pigment
presence of oils greatly increases its absorption being the most valuable microalgal compounds.
by the digestive system (Story, 2010). Haematococcus pluvialis strain is the richest
Lycopene is abundant in tomatoes, source of astaxanthin (Mezzomo and Ferreira,
watermelons, pink grapefruit, papaya and are 2016; Zaheer, 2017). This substance gives the
the most effective singlet oxygen scavenger in pinkish-red hue to the salmonids (salmons and
vitro (Sies and Sthal, 1995). Epidemiological trout), shrimps, lobsters and crabs and has an
studies concluded that diets rich in high important role in their immune-system and
lycopene foods may reduce risk of fertility. Nutritionally, astaxanthin is the most
cardiovascular diseases and different cancer powerful antioxidant in the nature, with role to
types (Boon C., 2010). scavenge efficiently free radicals from the
Torulene has one β-ionone ring and a longer human body (Koller et al., 2014).
polyene chain than that of β-carotene; it has 13 Similarly to lutein, it was proved to have a
double bonds and red color, in contrast to other positive role in many human health problems,
carotenoids produced by the microorganisms. such as: UV-light protection, anti-inflammatory
Torulene is found in red yeasts such as properties, support for eye health (helps
Rhodotorula glutinis, Sporobolomyces diabetic retinopathy, macular degeneration,

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eye strain and fatigue, and seeing in fine global carotenoids market is estimated to be
detail), immune-boosting effects (improves the valued at USD 1.53 Billion by 2021 and this is
ability of protective white blood cells to mainly due to the increasing number of health-
surround and destroy infecting organisms, conscious consumers and the rising demand for
especially fungi), prevention of different types natural coloration. The quality of food is
of cancer (colon and breast cancer) (Amaya et associated with color, flavor, texture, and odor,
al., 2014). humans considering color as the most relevant
Canthaxanthin is an orange-red oxycarotenoid aspect due to its appealing nature (Amaya et
found in crustaceans, fungi and algae, al., 2014).
particularly used in color feeding. If, today, carotenoids produced commercially
Canthaxanthin has an antioxidant activity and are mostly by total synthesis to meet the
enhances junctional communication between demands, intention is to growingly substitute
cells directly or through the formation of 4- these with natural ones, if possible obtained by
oxo-retinoic acid, which is also able to as mild as possible methods and processes
stimulate the retinoic acid receptor (Amaya et (Mezzomo and Ferreira, 2016).
al., 2014). Synthetic carotenoids
For carotenoid synthesis, one need to construct
SOURCES OF CAROTENOIDS FOR the polyene chain, meaning the hydrocarbon
FEED ADDITIVES skeleton conjugated by alternating double and
single bonds. For the specific location of the
As have been pointed out above, there has been double and single bonds, different reactions
a change lately in the perspective towards the have been developed along time, the first to be
ingredients of foods, what is considered healthy obtained being the β-carotene (Amaya et al.,
for humans and for the environment and what 2014).
should constitute the food of animals and birds. Out of economic and technological reasons,
Yet, researches are still conducted to efficiently although various carotenoids have been
obtain natural carotenoids and optimal feed synthesized at laboratory level, few of them
formulas for animals/birds. (Kaur and Shah, went to commercial scale, among which are the
2017). following: astaxanthin, astaxanthin-dimethyl-
According to the new study by di-succinate, as well as β-carotene, β-apo-8’-
reportsnreports.com (2016), the carotenoid carotenal, also canthaxanthin, citranaxanthin,
market is led by the feed segment. The demand and the ethyl ester of β-apo-8’-carotenoic acid.
for incorporation of carotenoids in animal feed For commercial use in animal nutrition, they
is growing due to consumers preference for are usually formulated as 10% concentrated
good quality and meat, meat products, and fish products as a powder or beadlet (Amaya et al.,
aspect. Carotenoids also increase the palata- 2014).
bility of feed, aquaculture being currently the Currently, carotenoids synthetic production is
fastest growing sector. It is known that the the most common manufacturing method. Over
farming of salmon and shrimp grew 95% of the feed market uses synthetic
exponentially in the last years and the trend is astaxanthin, mainly from major producers
ascendant, so the market of feed carotenoids, BASF (Germany) (Lucantin pink) and
especially of astaxanthin, expands rapidly Hoffman-La Roche (Switzerland) (Carrophyll
(Anonimous, 2010). pink), which contains only 25% of the biologi-
With the growth of world population and cally active stereoisomer found naturally. For
increased awareness and demand for better replacing, only in aquaculture feed additive, the
animal nutrition (in terms of quality and synthetic pigment with natural astaxanthin
quantity), carotenoid additives seem to be the produced by Haematococcus, the biomass of
solution (García-Chavarría and Lara-Flores, this microalga should be increased to 10 000
2013; Mohiseni, 2017). ton per year, the price of astaxanthin being too
According to global trends and forecasts high (Lorenz, 2000; Minyuk, 2008).
(https://www.marketsandmarkets.com/Market- The addition of synthetic carotenoids to the diet
Reports/carotenoid-market-158421566.html), of laying hens and broilers is a common

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practice, for improving yolk color (Santos- The majority of naturally occurring carotenoids
Bocanegra, 2004). Canthaxanthin is the are found in vegetables and fruits, dark green
preferred synthetic red xanthophyll in poultry leaves of plants and seeds, flowers and roots
farming available as Carophyll® Red (DSM (Amaya et al., 2014). Carotenoids found in
Nutritional Products, Switzerland) or plants differ in concentration depending on
Lucantin® Red (BASF, Germany). The plant varieties, degree of ripeness, time of
preferred yellow xanthophyll is β-apo-8´- harvest, growing and storage conditions, etc.
carotenoic acid ethyl ester available as (Zakynthinos et al., 2016).
Carophyll® Yellow and Lucantin® Yellow Feed ingredients such as corn, maize, tomato,
(Marounek, 2015). green beans and cabbage, pumpkins, prunes or
Synthetic carotenoids are technologically red pepper all are sources of carotenoids with
feasible and cost-effective. These are benefits for animals (Amaya et al., 2014;
standardized and more stable, i.e. apo-carotene- Zakynthinos et al., 2016; Mezzomo and
ester concentration in feed samples showed Ferreira, 2016,).
very little variation from expected values, Red pepper or paprika (Capsicum annuum) is
whereas it was 30% below expectations for reported to be a rich, abundant and inexpensive
marigold extracts. source of keto-carotenoids (capsanthin). It has
It has been determined that 1 ppm of apo- been studied as a fish feed additive but with a
carotene-ester is equivalent to about 2 ppm of lower efficacy in comparison to commercially
lutein-zeaxanthin from marigold. available astaxanthin and a less desirable
Canthaxanthin is the red carotenoid which coloration in comparison to canthaxanthin in
dominates pigmentation of broilers, whereas rainbow trout (Akhtar et al., 1999). In addition
paprika xanthophylls which require two to to pigmentation, paprika increased growth rate
three times the amount compared to and improved the reproductive performance in
canthaxanthin, receives little attention. yellow tail. Good results have also been
The stability of various red carotenoids reported for laying hens’ yolk pigmentation
products in premixes was checked after three (Yanar et al., 2016).
months of storage and recovery varied Marigold flower (Tagetes erecta) is rich in
considerably: 66% to 92% for canthaxanthin, lutein and zeaxanthin and it is efficient for
76% for citranaxanthin, and 39% for poultry skin and egg coloration. Lutein from
capsanthin from paprika, which shows that marigold is also used as a yellow coloration for
carotenoids are very unstable on their isomeric cultured marine fish yellow tail and red sea
forms. Another property is the transfer bream (Maoka T, 2011). As consumers
efficiency from feed to egg, which is better for associate color to nutritive value, healthiness,
the synthesized ones. Apo-carotene-ester has a freshness and taste, this may count as an
55% transfer efficiency to egg yolk, as interesting dietary alternative (García-
compared to only 17% for marigold Chavarría and Lara-Flores, 2013).
carotenoids and canthaxanthin an average Medicinal plants are known to be sources of
deposition rate of about 40% natural antioxidants used in treatments for
(www.poultryworld.net/Broilers/Nutrition/2012 human and animal diseases. Because of their
/8/The-effect-of-carotenoids-on-yolk-and-skin- fair price and strong effects, medicinal plants
pigmentation-WP010752W/). are rapidly becoming a safe alternative for
The consumer’s demand for “non-synthetic” antibiotics and chemical drugs in aquaculture.
colorants has driven the industry to identify and Furthermore, these can help increase growth as
develop natural sources of carotenoids to they maintain healthy intestinal microflora
replace chemical synthesis (Olesen, 2010). (Mohiseni, 2017).
Natural sources of carotenoids Some valuable natural supplements for
Plants pigmentation of chicken eggs, broilers and fish
are presented in the following Table 2.

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 Table 2. Natural animal feed supplements
Natural sources Main carotenoids Role as feed additive References
Carrots β-Carotene Increases yolk color parameters and carotenoid Isler, 1967
α-Carotene contents Hammershoj et al, 2010
Coloration of skin and flesh of food fish Weerakkody et al., 2016
Alfalfa (Medicago Lutein Pigmentation of broilers Amaya et al., 2014
sativa) Zeaxanthin Improve egg quality Varzaru et al., 2015
β-apo-8’-carotenal
β-apo-8’-carotenoic acid
Lycopene
Marigold flower Lutein Poultry skin and egg coloration Rajput et al, 2012
(Tagetes erecta) Zeaxanthin García-Chavarría and Lara-Flores,
2013
Yellow maize Lutein Boosts carotenoids in eggs and improve egg yolk Breithaupt, 2007
Cryptoxanthin, Zeaxanthin color
Natural sources Main carotenoids Role as feed additive References
Saffron (Crocus Crocetin Oxidative stability of shell eggs and liquid yolks Botsoglou et al, 2005
sativus) β-Carotene
Zeaxanthin
Seeds of Bixa orellana Bixin and decomposition products Animal nutrition Amaya et al., 2014
Paprika (red pepper) Capsanthin Pigmentation and weight gain of rainbow trout Yanar, 2016
(Capsicum annuum) Capsorubin Pigmentation of salmonids
Pigmentation of Pacific white shrimps Talebi et al., 2013
Yolk pigmentation in laying hens Arredondo-
Weight gain in broilers Figueroa et al., 2003
Vicente et al., 2007
Galib et al., 2011

There are some problems regarding carotenoids 70% of the raw weights of the catch are
production from plant origin, due to seasonal processing discards” reducing at the same time
and geographic variability that cannot be an environmental problem (Shahidi et al.,
controlled. A better option, regarding yields 1998). Lobsters, crabs and shrimps, together
and costs, is the microbial production of with their processing waste are sources of
carotenoids on agro-industrial wastes which are astaxanthin (Mezzomo et al., 2016).
cheap substrates (Mata-Gómez, 2014). Supply of marine animal based natural
Crustacea and marine animals carotenoids are limited because of declining
Marine animals accumulate various carotenoids trend in catches of crustaceans from marine
from foods (bacteria, algae, other animals) and resources. These sources of carotenoids are
modify them through metabolic reactions very expensive and thus aquaculture feed
(oxidation, reduction, translation of double production involves high production costs
bonds, oxidative cleavage of double bonds, (García-Chavarría and Lara-Flores, 2013).
cleavage of epoxy bonds). Many of these Microbial sources of carotenoids
carotenoids are metabolites of β-carotene, Interest in microbial sources of carotenoids was
fucoxanthin, peridinin, diatoxanthin, renewed due to consumers opinions against
alloxanthin, and astaxanthin (Maoka, 2011). synthetic additives. Types of carotenogenic
Astaxanthin is distributed in both marine and microorganisms, like bacteria and fungi, can be
fresh water fish. Cyprinidae fishes synthesize stimulated to produce cost-effective
(3S,3′S)-astaxanthin from zeaxanthin by carotenoids in response to environmental stress
oxidative metabolic conversion. Perciformes (light, temperature), chemical compounds or
and Salmonidae fish cannot synthesize modification of metabolic pathway using
astaxanthin from other carotenoids (Matsuno, recombinant DNA technology (Bhosale,
2001). Astaxanthin present in these fishes 2004). Advances have been made through
originates from crustacean sources (Atlantic genetic manipulation of some non-
krill, crayfish meal, crab meal) used in carotenogenic microbes such as Escherichia
aquaculture feed formulation as additive. coli, Saccharomyces cerevisiae, Candida utilis
Crustacean by-products represent an “attractive in terms of carotenoids production (Das et al.,
ingredient for industrialization, since around 2007).
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Several bacteria have biotechnological Mycobacterium brevicaie, Mycobacterium
potential for the production of carotenoids. lacticola, Rhodobacter sphaeroides,
Some thermophilic halophilic bacteria such as Rhodococcus maris, Streptomyces
Halococcus morrhuae and Halobacterium chrestomyceticus (Dannert, 2000).
salinarum develop red and orange colonies, Some examples of carotenoid-producing
bacterioruberin synthetized by H. salinarum microorganisms are presented in Table 3.
being the most found carotenoid (Asker and Practical obstacles still need to be overcome for
Ohta, 1999). Haloferax alexandrines has good their commercial exploitation, though
industrial perspective for the production of developments in recent years have proven that,
canthaxanthin (Asker and Ohta, 2002) and the in both farmed fish and poultry, carotenoids
marine bacterium Flavobacterium sp. produces like astaxanthin, canthaxanthin, β-carotene,
zeaxanthin, being considered one of the best zeaxanthin and lycopene from bacterial sources
microbial sources of zeaxanthin (Masetto et al., impart color to skin, flesh and eggs (Bhosale,
2001). Other bacteria with ability to synthesize 2004).
carotenoids are Agrobacterium aurantiacum,

Table 3. Microorganisms sources of carotenoids and their application as feed additives

Bacteria and Yeasts Carotenoids/Activity Application as animal feed additives References
Mycobacterium lacticola Astaxanthin/ antioxidant, Fish feeds Kushwaha et al., 2014
photo- protectant
Paracoccus carotinifaciens Astaxanthin/ Coloration of farm raised salmon and Bories, 2007
(commercially sold as Panaferd) pigmentation trout
Spongiococcum excentricum Lutein/antioxidant Poultry feeds Kushwaha et al., 2014
Flavobacterium sp. Zeaxanthin/ Additive in poultry feed to increase Alcantara and Sanchez, 1999
pigmentation yellow color of animal’s skin and egg Masetto et al., 2001
yolk Bhosale, 2004
Fish feed
Haloferax alexandrines Canthaxanthin/ Feed in salmon farming to guarantee Asker and Ohta, 2002
pigmentation the flesh color of fish
Dietzia natronolimnaea HS-1 Canthaxanthin/ Egg yolk pigmentation Esfahani-Mashhour et al., 2009
pigmentation, antioxidant Gharibzahedi et al., 2012
Phaffia rhodozyma Astaxanthin/ Pigmentation of salmon, trout, and red Maoka, 2011
pigmentation sea bream (Pagrus sp.) Kushwaha et al., 2014
Pigment source for egg yolk of laying
hens
Blakeslea trispora β-carotene/growth Chickens for fattening Sales et al., 2008
performance Shrimp feed
Feed for dairy cows
Rhodotorula spp. β-carotene Nutrition of laying hens Bhosale P., 2004
Torulene Kushwaha et al., 2014
Torularhodin/
antioxidant, precursor of
vitamin A
Xanthophyllomyces dendrorhous Astaxanthin Feed supplement for salmons, crabs, Bhosale P, 2004
shrimps, chickens and egg production Kushwaha et al., 2014
Mushrooms
Cantharellus cinnabarinus Canthaxanthin Poultry feeds and fish feeds Kushwaha et al., 2014

Microbial sources are an environmental-friendly researched and commercialized both for

method for the production of carotenoids, which nutritional supplements for humans and as
meet the increasing demand of these natural animal feed additives to replace synthetic
products (Das, 2007). components (Priyadarshani and Rath, 2012).
Microalgae, rich source of carbohydrates, Microalgae play an important role in the high-
protein, enzymes and fiber, have been a major level nutrition for both aquaculture and farm
source of food for humans in Asian countries. animals. The most famous sources of
They have been naturally taken over, microalgae, which accumulate carotenoids in
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their biomass, are Chlorella, Chlamydomonas, sources of natural pigments (β-carotene, α-
Dunaliella, Muriellopsis and Haematococcus carotene, β-cryptoxanthin and zeaxanthin,
spp, offering economical alternatives to respectively) for the culture of black tiger
chemical synthesis (Bhosale and Berstein, prawns (Penaeus monodon), salmonid fish and
2005). ornamental fish (Priyadarshani and Rath,
Carotenoids, even at levels of parts per million, 2012). Zeaxanthin from Spirulina is used as a
are strong dyes, especially canthaxanthin, red coloration for goldfish and ornamental
astaxanthin and lutein from Chlorella, which carp. Astaxanthin became the most important
have been used as pigments and included as carotenoid in salmons and rainbow trout with
ingredients in feed for salmonid fish, trout, and the development of salmonid farming.
poultry to enhance the reddish color of fish and Astaxanthin can be incorporated in feeds up to
egg yolk yellowish color (Plaza et al., 2009). 100 mg/ kg and canthaxanthin up to 25 mg/kg
Broiler chickens fed with algae have yellow (high accumulation in humans can be toxic).
skin and shanks, and the egg yolk is darker Animal feed is the largest field of commercial
indicating higher carotenoids content (Becker, application of microalgae carotenoids due to
2004). Microalgae like Chlorella vulgaris and the importance of astaxanthin and
Haematococcus pluvialis synthetize large canthaxanthin in aquaculture (Britton, 2004).
amount of astaxanthin (Yin, 2013; Kim, 2016) Bellow given is Table 4, including the sources
and Dunaliella salina, Spirulina maxima and S. of carotenoids from microalgae with
platensis are biotechnologically relevant, as application as feed additives.

Table 4. Microalgae sources of carotenoids

Algae and cyanobacteria Carotenoid Application as feed additives References
Haematococcus pluvialis Astaxanthin Animal feed additives Priyadarshani and Rath, 2012
(commercially sold as Aquaculture Mezzomo and Ferreira, 2016
NaturRose) Used for culture of prawns, salmonid García-Chavarría, Lara-Flores, 2013
fish and for ornamental and tropical Bhosale, 2004
fish Amaya et al., 2014
Dunaliella sp. β-carotene Animal feed additives Priyadarshani and Rath, 2012
Used as natural food coloring agent in Mezzomo and Ferreira, 2016
aquaculture feed industry García-Chavarría, Lara-Flores, 2013
Bhosale, 2004
Amaya et al., 2014
Chlorella vulgaris Lutein Used in diets of rainbow trout yielding Priyadarshani and Rath, 2012
Astaxanthin both muscle and skin pigmentation Mezzomo and Ferreira, 2016
Fucoxanthin effects García-Chavarría M., Lara-Flores M, 2013
Animal feed additives
Chlorella protothecoides Lutein The reddish color of salmonid fish and Guedes et al., 2011
yellowish color of egg yolk Mezzomo and Ferreira, 2016
Algae and cyanobacteria Carotenoid Application as feed additives References
Chlorella zofingiensis Lutein Enhances the reddish color of salmonid Guedes et al., 2011

Muriellopsis sp. Lutein Poultry farming and/or aquaculture Blanco et al., 2006
Scenedesmus obliquus Lutein Fish feed Mezzomo and Ferreira, 2016
Tetraselmis sp Lutein Aquaculture and larval feeds Priyadarshani and Rath, 2012
Nannochloropsis oculata Astaxanthin Aquaculture & larval feeds Priyadarshani and Rath, 2012
Hen supplements - increases egg Zaheer, 2017
nutritional value Gładkowski et al, 2011
Spirulina sp β-carotene Animal feed additives Indira Priyadarshani and Biswajit Rath, 2012

The market demand for pigments natural The European legislation specifies that before
sources has promoted large-scale cultivation of their use in EU, carotenoids have to be
microalgae for biosynthesis of such compounds subjected to an authorization process.
(well-established are β-carotene and A general overview of the current legal
astaxanthin), so significant decreases in situation of the different carotenoids used as
production costs are expected in coming years. feed additives in compound feed in European
81
Union is shown in Table 5 (Vincent et al., (Pérez-Gálvez, 2003; Baldo, 2011; Amaya et
2017), with added data regarding what al., 2014; Rodriguez-Amaya, 2015; Mezzomo
carotenoids are totally synthesized and which and Ferreira, 2016).
are extracted or biosynthesized come from

Table 5. Carotenoids used in the feed industry as feed additives in the European Union
Carotenoids feed Target animal Total Extracted Biosynthesized EURL-FA evaluation
additive synthesized report

Astaxanthin Fish (salmon, trout) Crustaceans Phaffia rhodozyma EURL-FA Feed

Pink shrimp Haematococcus pluvialis Additives (2014)
(Penaeus Chlorella vulgaris
paulensis)
processing
waste
Astaxanthin Fish X EURL-FA Feed
Ornamental fish Additives (2010a)
Crustaceans
Astaxanthin Salmon and trout X EURL-FA Feed
dimethyldisuccinate1 Additives (2007a)
β-apo-8’-carotenoic Poultry X EURL-FA Feed
acid ethyl ester Additives (2011c)
(apoester)2
Astaxanthin, Salmon and trout Red carotenoid-rich EURL-FA Feed
adonirubin, Paracoccus Additives (2007b)
canthaxanthin carotinifaciens
Canthaxanthin2 Chickens for fattening and minor X Chlorella vulgaris EURL-FA Feed
poultry species for fattening; Additives (2011a)
Laying poultry and poultry reared
for laying;
Ornamental fish and ornamental
birds;
Ornamental breeder hens
Capsanthin3 Poultry Red peppers EURL-FA Feed
Additives (2012b)
Citranaxanthin2 Poultry X EURL-FA Feed
Additives (2016)
Lutein3 Poultry Marigold EURL-FA Feed
(Tagetes erecta Additives (2012a)
and Tagetes
patula)
Rose fruit
(Rosa canina)
Carrot
β-carotene (solid All animal species X Chlorella vulgaris EURL-FA Feed
form) Dunaliella salina Additives (2010b)
Arthrospira maxima
Zeaxanthin3 Poultry Marigold EURL-FA Feed
(Tagetes erecta Additives (2012a)
and Tagetes
patula)
Paprika
(Capsicum
annuum)
1
formulated in an organic matrix
2
formulation containing the active substance and other ingredients
3
formulation containing a natural source of the active substance and other ingredients
EURL-FA - EU Reference Laboratory for Feed Additives

In the EU, carotenoids used as feed additives Union is currently limited to 80 ppm (mg/kg),
are mainly authorised under the category excepted canthaxanthin, whose limit is 8 ppm,
„sensory additives”, as colorants (Vincente et because excessive exposure in humans can lead
al., 2017). The addition of natural and synthetic to the development of precipitated crystals in
carotenoids in layer feeds in the European the retina (Baker, 2001).
82
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