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September | October 2014

Yeast cell wall and the immuno stimulation

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FEATURE

Yeast
cell wall
and the
immuno
stimulation

he world aquaculture activities have shown a rapid increase is the primary defence mechanism against infectious disease. A local
in production (17.8 percent per year) since 1997 (FAO, immune response in the intestinal mucosa is generated in different steps.
2007, Mohamed et al. 2010). With intensification of aqua- At first macrophages and other phagocytic cells, like dendritic cells, take
culture production, the spreading of disease has been the up the pathogen (nonspecific immunity), and destroy them with enzymes
main problem in the fish farming industry. Up until now, antibiotics and O-radicals, and process it for presentation to local T cells. Phagocytic
were used as treatment for fish
disease management and also in Table 1. Total count of bacteria, immunity items and proximate composition of the common carp fingerlings
order to offer a better growth fed diets containing different levels of ImmunoWall for 8 weeks.
performance and feed efficiency.
0.0 g/kg 0.5 g/kg 1.0 g/kg
1.5 g/kg
2.5 g/kg
As in the other animal species, the
Parameters
widespread usage of antibiotics as a
Total counts of bacteria (Log
9.18
7.78
7.50
7.30
6.96
feed additive in aquaculture has been
cfu/g intestine)
0.23
0.26
0.44
0.18
0.17
under intensive scrutiny, as there
Immunity items
are growing concerns over the risk
Albumin (g/l)
11.5
10.70
11.3
11.5
12.1
associated with the transmission of
0.4
0.7
0.5
0.5
0.8
resistant bacteria from aquaculture
Globulin (g/l)
13.09
12.7
13.6
14.6
15.6
environments to humans, and also
0.9
0.1
0.9
0.7
0.6
the risk associated with the introAlbumin:Globulin ratio
0.82
0.84
0.83
0.78
0.77
duction in the human environment
0.08
0.12
0.09
0.07
0.08
of nonpathogenic bacteria, containLeucocyte(x10 mm)
2.01
2.28
2.25
2.24
2.15
ing antimicrobial resistance genes,
0.04
0.01
0.02
0.03
0.02
and the subsequent transfer of such
Total protein (g/l)
25.4
23.4
24.9
26.1
27.7
genes to human pathogens.
On January 2006, the European
Union ratified a ban on the use
of all sub-therapeutic antibiotics as
growth-promoting agents in animal
production. Concomitant with the
ban of antibiotic growth promoters
(AGP) new strategies in feeding and
health management in fish aquaculture practice have received much
attention. The research of prebiotics
in fish nutrition is increasing with the
demand by consumers for safe products produced according the sustainable practices (Denev et al, 2009).

Aquaculture And
Protection Mechanism

The immune system of the fish

0.55

0.35

0.40

0.25

0.15

Glucose (mg/l)

1196
2.51

1201
3.69

1200
3.60

1199
3.83

1195
3.51

Haematocrit (%)

27.7
0.72

27.9
0.35

26.9
0.46

26.6
0.37

26.4
0.65

Proximatencomposition (g/kg)

Initial
values

Moisture

746.66
6.43

715.33
4.37

709.07
2.99

708.72
3.11

707.51
5.12

703.93
3.64

Protein

132.3
3.6

145.18
1.21

146.57
1.72

150.79
1.68

153.21
2.24

156.40
1.96

Lipid

82.1
1.91

95.54
1.81

95.81
1.18

96.72
1.19

98.72
1.47

98.43
1.65

Ash

26.5
0.53

27.45
0.66

28.60
0.50

28.04
3.66

28.73
0.80

28.70
0.60

Energy

6.53
0.7

7.42
0.10

7.52
0.04

7.58
0.06

7.65
0.11

7.73
0.11

P:E ratio

20.27
0.56

19.55
0.24

19.46
0.11

19.88
0.35

20.0
0.46

20.22
0.09

18 | INTERNATIONAL AQUAFEED | September-October 2014

FEATURE
cells also secrete pro-inflammatory cytokines that stimulate the activity
and proliferation of those T and B cells that are most fit to attack the
type of pathogen that was presented by the phagocytic cell (specific
immunity) (Ghent, 2011).
Immunity in fish has an important role against pathogen agents. Fish
immune systems share some similarities to mammals. Fish, however,
rely more on non-specific defence mechanisms than mammals do. The
non-specific immune system of fish consists of several key humoral and
cellular components that provide innate protection against infection,
regardless of the pathogen type. Several studies have demonstrated
that immune-stimulants are very beneficial to fish immune system
(Denev et al., 2009)
Various mechanisms have been proposed to explain the specific
action of immune stimulants in fish, such as selective stimulation of
beneficial microbiota, improvement of immune functions, disease
resistance, survival, growth performance and feed efficiency.

Beta Glucansas Yeast Cell Wall Component

They have been studied as immune stimulants in numerous

vertebrate and invertebrate species. The evaluation of cellular and
humoral immune parameters was established at phenotypic level by
the phagocytic activity of head kidney macrophages and quantification
of inflammatory cytokines in spleen and gill tissues by indirect ELISA.
Innate immune response, mediated by lysozyme, was reported in
several fish species.
Prebiotics are non digestible functional fibres that beneficially affect
the host by selectively stimulating the growth of and/or activating the
metabolism of one or a limited number of health-promoting bacteria
in the intestinal tract, thus improving the hosts intestinal. Beta-glucan
is a particulate carbohydrate that consists of glucose and mannose
and is the main constituent of the yeast cell wall (Schimitt et al., 2013).
Several studies confirming the potent immune stimulatory properties
of beta-glucans in many fresh and seawater fish species documenting
the effects themon the pathogen resistance, protection, survival, and
fish specific humoral immunity have been published. Strong degranulation of neutrophils was observed in fathead minnow after dietary
administration of beta-glucans. The lysozyme activity increased after
4 weeks of feeding the whole yeast cells at a dose rate 10g/kg ration
to gilthead seabream. Enhanced phagocytic activity, respiratory burst
activity, and bactericidal action in the head kidney macrophage were
observed in the groups that received dietary immune stimulants.

Yeast Cell Wall - Immunity and Resistance

to AeromonasHydrophila Infection

Several studies with ImmunoWall (ICC) were carried out in Sari

Agriculture University (Ebrahimi, 2011). ImmunoWall is a natural
product containing various stimulating compounds such beta-glucan,
complex carbohydrates, and MOS, glucomannoprotein complexes,
which have been used as feed additives in animal production. Both
are components of the yeast cell wall (Saccharomyces cerevisiae).
Appropriate or low levels of beta glucans were stated to be efficient
stimulators of non-specific immune functions in fish.

Material and Methods

The fingerlings were adopted for 2 weeks and then reared in

triplicate groups in 250-l tanks (n = 15 per tank with average initial
weights of 11.12 0.55 g), and divided into five groups that received
experimental diets (isonitrogenous and isoenergetic) containing different levels of ImmunoWall (0, 0.5, 1, 1.5 and 2.5 g prebiotic/ kg diet)
thrice a day (until apparent satiation) for 8 weeks.

Experimental Results

Both feed efficiency ratio and protein efficiency ratio significantly

(p < 0.05) increased with increasing ImmunoWall levels from 0.5
to 1.5 g/kg diet. The highest protein content (p < 0.05) was found
September-October 2014 | INTERNATIONAL AQUAFEED | 19

FEATURE
in the fish fed a diet containing 2.5 g/kg prebiotic. Haematological
parameters and plasma total protein concentration were also significantly higher (p < 0.05) in the fingerlings fed diets containing 1.5 and
2.5 g/kg prebiotic in relation to the control.
The control fish contained the highest mean of total bacterial
counts. The lowest mean (p < 0.05) of total bacterial counts were
observed in the fish fed the diet containing 2.5 g/kg ImmunoWall.
The present study reveals that a dietary ImmunoWall supplementation from 1 to 1.5 g/kg is capable to improve the feed efficiency and
growth performance of C. carpio fingerlings as well as their resistance
to A. hydrophila infection and the ImmunoWall levels from 0 to 2.5
g/kg diet revealed no mortality throughout the experiment.

Discussion

It has recently been shown by Jafar Nodeh (2010) that supplementing the diet of the Persian sturgeon (Acipenser persicus) fingerlings with
the commercial prebiotic ImmunoWall considerably modified the
intestinal microflora. The author also reported that Lactobacillus (lactic
acid bacteria) population in the intestine of the fish fed a diet containing
2 g prebiotic/kg, was higher than that of other supplemented groups,
while total counts of bacteria steadily declined with an increase in the
prebiotic levels up to 2 g/kg diet (p < 0.05). Furthermore, it has been
confirmed, in a number of studies, that lactic acid bacteria as one of the
common probiotics have beneficial effects on the improvement of host
nutrition through the production of supplemental digestive enzymes;
they also produce bacteriocins that inhibit specific fish pathogens.
It seems that changes in villi morphology and density in the common
carp fingerlings fed with the prebiotic ImmunoWall are probably one
of the remarkable factors involving in the increased potential nutrient
capture thereby increased feed efficiency.
The increase in WBC (White blood cells) count might be due
to stress suffered by fish as a result of daily feeding on beta-glucan.

Harikrishnan et al. (2003) also reported increased WBC counts in

C. carpio after herbal treatment with Azardicha indica. The observed
increases in the leucocyte and total protein levels as well as lower
mortalities resulting from the pathogenic A. hydrophila infection
appear to be signs of enhanced health status of the prebiotic-fed fish.
Moreover, high concentrations of serum proteins including humoral
elements of the non-specific immune system are likely to be results
of an enhancement in the non-specific immune response of the fish.
The improved health condition in the C. carpio fingerlings is probably
due to the beta-glucan and MOS components of the ImmunoWall.
Glucans can modulate the activity of phagocytes and other components of the innate immune system in fish (Robertsen et al., 1994).
The administration of glucans in the diet of fish has been represented
to improve the respiratory burst activity, phagocytosis, lysozyme value
and resistance vs. pathogens in some cultured fish (Ogier de Maulny
et al., 1996; Li and Gatlin, 2004). Rodriguez et al. (2007) found that
b-1, 3/1 6- glucans would modify the immune response of P. vannamei juveniles, influencing the white spot syndrome virus (WSSV)
prevalence and shrimp survival in ponds. Li and Gatlin (2004) observed
that hybrid striped bass fed glucan and/or chitin from brewers yeast
showed extracellular superoxide anion production of head kidney
macrophages. This prebiotic supplement also improves the fish resistance to A. hydrophila infection, thereby acting as a potent immune
stimulant. Notwithstanding, extra dosage of ImmunoWall seems to
hinder enhancements of feed utilization and growth of the fingerlings.

Conclusion

Based on the findings of this study, supplementation of

ImmunoWallat levels of 11.5 g/kg was beneficial on immune
system, increasing the performance and the survival of C. carpio
fingerlings.
Full citations and references available on request

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