Nowadays, all animal production is concerned with vaccination. This is an essential technique for the protection of livestock health which, however, entails significant costs for stock breeders. Maximizing the efficiency and profitability of prophylactic vaccination strategies is therefore a major stake. To achieve this, new avenues are constantly explored. One of these concerns is the use of new molecules extracted from seaweeds to help optimize the stimulation of the natural defences of the body and its response to vaccination strategies.

1. September | October 2013
Marine algal polysaccharides: a new option
for immune stimulation
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3. N
owadays, all animal production
is concerned with vaccination.
This is an essential technique
for the protection of livestock
health which, however, entails significant
costs for stock breeders. Maximizing the
efficiency and profitability of prophylactic
vaccination strategies is therefore a major
stake. To achieve this, new avenues are
constantly explored. One of these concerns
is the use of new molecules extracted from
seaweeds to help optimize the stimulation
of the natural defences of the body and its
response to vaccination strategies.
Innate immunity
The body’s response to the aggression of a
pathogen is based on two types of immunity.
They are the innate immune response and the
adaptive response.
The innate response is the first line of
defence against pathogens. It is activated
immediately and acts very quickly. This
immune response can be found in all ani-
mals. It will be the same whenever the
body encounters that pathogen. However,
the body does not retain a memory of the
infectious agent. The mechanism of action of
this type of immunity consists in recognizing
the molecular patterns shared by numerous
pathogens, which are essentially represented
by membrane fractions (glycocalyx).
The various elements that contribute to
the innate immune response are the following:
• Physical barrier (mucous membrane,
skin, mucus, villi etc)
• Phagocytic cells, such as the macrophages
• Natural killer (NK) cells
• Certain cytokines, which deliver signals
warning the body of a danger
• Complement system
• Toll-like receptors (TLR), a family of
membrane receptors only discovered
recently. They control the expression
of molecules that fight against infectious
agents (directly or indirectly, via effector
cells, and by recruiting the activation of
the adaptive immune system).
The elements associated with the innate
immune response can act on the pathogen
directly or indirectly, by producing effector
cells (cytokines etc). The latter will subse-
quently trigger the adaptive immunity by
activating the T and B cells.
Adaptive immunity
Unlike the innate response, the acquired or
adaptive response occurs in vertebrates only.
During the first encounter with a given patho-
gen (primary infection), it acts as the body’s
second line of defence. Its activation takes
some time - known as latency. However, this
response system memorizes the pathogens
it encounters and when the body is again
exposed to them the latency is much shorter
and the immune system reacts to the aggres-
sion almost immediately. Adaptive immunity
is specific: it recognises the molecular patterns
of the already encountered pathogens.
The various elements that contribute to
the adaptive immune response are the fol-
lowing:
• T cells
• B cells
• Antibodies
• Ig, TCR, CTL, antibody (AB)-producing
plasma cells + coupled aid of the innate
immunity effectors
Seaweeds: a new source of
active elements to stimulate
the immune system
In recent years more and more publica-
tions have brought to the forefront the
relevance of seaweeds in numerous biological
applications, particularly to immune mecha-
nisms, taking special interest in some of their
components, namely the sulfated polysac-
Marine algal polysaccharides:
a new option for immune stimulation
18 | InternatIOnal AquAFeed | September-October 2013
FEATURE
NK : Natural Killer
PRR : Pattern Recognition Receptor
CMH : Complexe Majeur d’histocompatibilité
TCP : T Cell Receptor

5. charides. These are complex carbohydrates
which do not occur in terrestrial plants. They
are supposed to influence the immune system
by a vast number of still poorly understood
pathways.
Polysaccharides represent a structurally
diverse class of macromolecules which are
relatively widespread in nature. There are
simple and complex forms of polysachharides.
Unlike proteins and nucleic acids, polysac-
charides contain repetitive structural features
which are chains of monosaccharide residues
joined together by glycosidic bonds.
Thus, they form polymer (-type) structures
represented in the form of chains that may be
homogenous (homopolysaccharides) or not
(heteropolysaccharides). The simple forms
are the homopolysac-
charides composed of
a single type of sugar,
linked in an essentially
linear manner (starch,
glycogen, cellulose for
example). They are
essentially structural
compounds or mecha-
nisms of energy storage
in an easily releasable
form. Their structure
may become more
complex owing to their capacity to
establish links at various levels of each
elementary unit, allowing thus the
development of branching structures
in the three dimensions. These are the
branched heteropolysaccharides.
Structural variability and
biological potentialities
The nucleotides in nucleic acids and
the amino acids in proteins can inter-
connect in only one way, while the
monosaccharide units in oligosaccha-
rides and polysaccharides can intercon-
nect at several points to form a wide
variety of linear or branched structures
(Sharon and Lis 1993). For instance,
the number of possible permutations for
four different sugar monomers can attain
September-October 2013 | InternatIOnal AquAFeed | 19
FEATURE
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Innate immunity / acquired immunity: two complementary and cooperative systems
Innate immunity adaptive or acquired immunity
Chronology
Primary infection
Quick response: first
barrier against pathogens
Second line of defence : latency
(about 7 days)
repeated infections
Identical to the primary
response
Immune memory => latency
close to zero
Specificity non-specific response Specific response (Ig and tCr)
recognized molecular patterns
Invariable and common to
numerous pathogens
Specific to the infectious agent
Cellular and molecular effectors
Complement, phagocytic
cells and certain cytokines
Ctl (cytotoxic t cells) and anti-
body producing plasma cells,
with the help of innate effectors
tCr : t Cell receptor - Ig : Immuno Globulin - Ctl : Cytotoxic t lymphocyte or Killer t cells
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7. up to 35,560 unique tetrasaccharides, while
four amino acids can form only 24 different
permutations (Hodgson 1991).
This explains the fact that, among macro-
molecules, polysaccharides provide the high-
est capacity for carrying biological informa-
tion, as they have the greatest potential for
structural variability. In addition, one of the
particularities that numerous marine polysac-
charides possess is their polyanionic character,
which confers them a high chemical reactivity.
Of these anionic polysaccharides, the
majority of those which occur macroalgae
are sulfated polysaccharides: galactan (agar,
carraghenans), ulvans, fucans.
The ulvans, for example, the water-soluble
polysaccharides found in green seaweed of
the order Ulvales (Ulva and Enteromorpha),
have sulfate, rhamnose, xylose and iduronic
and glucuronic acids as their main constitu-
ents (Lahaye and Ray 1996) (Percival and
McDowell 1967).
Ulvan structure shows great complexity
and variability as evidenced by the numerous
oligosaccharide repeating structural patterns
identified (Lahaye and Robic 2007). The main
repeating disaccharide units reported are of
ulvanobiouronic acid 3-sulfate type, containing
either glucuronic or iduronic acid. In addition,
a few repeating patterns can be found that
contain sulfated xylose replacing uronic acid
or glucuronic acid on the O-2 binding/link of
the rhamnose-3-sulfate units (Lahaye and Ray
1996) (Lahaye et al. 1997).
Interests
This huge variability in the polysaccharide
structure provides the flexibility required for
exact regulatory mechanisms in different cell-
cell interactions in higher organisms.
Sulfation in particular seems to be con-
ducive to various biological activities noted
in polysaccharides extracted from marine
macroalgae.
Marine sulfated polysaccharides:
their role and effect on immunity
Sulfated polysaccharides, which are wide-
spread in macroalgae, have been shown
to possess anti-infectious (Cumashi et al.
2007) (Witvrouw and De Clercq 1997) (anti-
viral, anti-bacterial, anti-tumoral), antioxidant
(Wang et al. 2010) (de Souza et al. 2007) and
anti-thrombotic (Mao et al. 2006) activities,
as well as immune-modulating (Leiro et al.
2007) activities that might find relevance in
stimulating the immune response or in con-
trolling the activity of immune cells in order to
mitigate negative effects such as inflammation
(Chen et al. 2008) One of the pathways of
marine sulfated polysaccharides, which has
been emphasized recently, is their role in the
activation of TLR.
Indeed, more and more studies are demon-
strating that marine algal polysaccharides can influ-
ence the innate immune response by binding to
recognition receptors called Pattern Recognition
Receptorsc (PRR), such as the mannose recep-
tors or TLRs of phagocytic cells, including and
especially macrophages (Chen et al. 2008). TLRs
are transmembrane proteins which detect invad-
ing pathogens by binding to ancestral molecules
of microbial origin called Pathogen-Associated
Molecular Patterns (PAMPs).
The PAMPs contact at TRL level triggers a
cascade of responses resulting in the expres-
sion of inflammatory response genes. In mam-
mals, these recently identified receptors have
been numbered from 1 to 11 (TLR1-TLR11).
On contact with their
respective PAMPs,
TLR specifically acti-
vate a signaling path-
way leading to the
activation of NF-kB
(Nuclear Factor-
kappa B) and AP1
(ActivatorProtein
1) transcription fac-
tors regulating the
expression of inflam-
matory cytokines
such as TNFα, IL-1
or IL-6.
It therefore now
20 | InternatIOnal AquAFeed | September-October 2013
FEATURE
Structure of the 4 main repeating ulvan
patterns of Ulva lactuca
Ulvanobiouronic acid A. [→ 4)-β-D-
GlcA-(1 → 4)-α-L-Rha3S-(1 →] n;
Ulvanobiouronic acid B. [→ 4) - α-L-
IdoA-(1 → 4)-α-L-Rha3S-(1 →] n
Ulvanobiose A. [→ 4)-β-D- Xyl -(1 →
4)-α-L-Rha3S-(1 →] n;
Ulvanobiose B. [→ 4)-β-D- Xyl 2S-(1 →
4)-α-L-Rha3S-(1 →] n;
Classification of Marine Sulfated
Polysaccharides (MSP)
Branched sulfated
heteropolysaccharide

8. appears that TLR play a key role in the
adaptive immune response, but the signals
produced by their activation lead to the
activation of numerous other cells and func-
tions of the immune system, which makes
them essential elements of both the innate
immune mechanisms and of adaptive immu-
nity.
The activity of some sulfated algal polysac-
charides as TLR activating agents might be
the result of a certain structural similarity
between these marine polysaccharides and
bacterial lipopolysaccharides (LPS). Bacterial
LPSs are indeed a type of structure occurring
at the surface of their external membrane
and recognized as bacteria-specific recognition
elements. In particular, bacterial LPS in mam-
mals are shown to be specifically recognised
by TLR4.
Possible applications
in animal health
In conclusion, seaweeds appear to contain
sugars in the form of polysaccharides, some of
which - sulfated polysaccharides - are complex
polyanionic structures which possess various
biological properties. A vast number of studies
have already evidenced the effects of some of
these sulfated polysaccharides, particularly the
fucoidans, the carraghenans and the ulvans, on
certain mechanisms of inflammatory response
and on immunity.
The identification and selection of these
polysaccharides extracted from suitable
macroalgae makes it possible to envisage
the use of these molecules as agents for
the stimulation of the various mechanisms
associated with the body defence and, in
particular, of the innate immunity mecha-
nisms.
Within the framework of the potential
applications in the fields of animal breeding
and animal health two non-exclusive strategies
can be proposed:
Regular sequential intakes for a general
stimulation of the body’s state of defence:
• With a regular intake not connected
with vaccination, they allow the
strengthen-
ing of the
body’s state
of defence.
Repeated use
allows the
development
of a ‘basic’
immune sys-
tem and the
boosting of
the state of
defence of the
innate system.
The use of
polysaccha-
rides upstream
or downstream
of a prophylactic
programme may
be an asset in
enhancing the
level of immune
protection of
an individual or
group of indi-
viduals within a
livestock and in
contributing to
a better control
of the infectious
pressure on
the livestock,
preventing the
appearance of
recurrent infec-
tious pathologies.
Targeted intakes
within the framework
of a vaccination pro-
gramme:
• As part of a
vaccination pro-
gramme, they
would enhance
the vaccine
protection. This
would definitely
provide the pos-
sibility to improve
the intake and
persistence of the
vaccine and there-
by to improve
the technical and
economic per-
formance of vac-
cine prophylactic
programmes.
More InforMatIon:
Website:www.olmix.com
September-october 2013 | InternatIonal AquAFeed | 21
FEATURE
Sulfated
polysaccharide
Cytokine/Chemokine
production

9. www.aquafeed.co.uk
LINKS
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• Visit the International Aquafeed website
• Contact the International Aquafeed Team
• Subscribe to International Aquafeed
The potential of
microalgae meals
– in compound feeds for aquaculture
Understanding ammonia
in aquaculture ponds
Volume 16 Issue 5 2013 - sePTemBeR | oCToBeR
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