yeast cell wall and the immuno stimulation

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September | October 2014Yeast cell wall and the immuno stimulation

The International magazine for the aquaculture feed industry

International Aquafeed is published six times a year by Perendale Publishers Ltd of the United Kingdom. All data is published in good faith, based on information received, and while every care is taken to prevent inaccuracies,the publishers accept no liability for any errors or omissions or for the consequences of action taken on the basis ofinformation published.©Copyright 2014 Perendale Publishers Ltd. All rights reserved. No part of this publication may be reproduced in any formor by any means without prior permission of the copyright owner. Printed by Perendale Publishers Ltd. ISSN: 1464-0058

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The world aquaculture activities have shown a rapid increase

in production (17.8 percent per year) since 1997 (FAO,

2007, Mohamed et al. 2010). With intensification of aqua-

culture production, the spreading of disease has been the

main problem in the fish farming industry. Up until now, antibiotics

were used as treatment for fish

disease management and also in

order to offer a better growth

performance and feed efficiency.

As in the other animal species, the

widespread usage of antibiotics as a

feed additive in aquaculture has beenunder intensive scrutiny, as there

are growing concerns over the risk

associated with the transmission of

resistant bacteria from aquaculture

environments to humans, and also

the risk associated with the intro-

duction in the human environment

of nonpathogenic bacteria, contain-

ing antimicrobial resistance genes,

and the subsequent transfer of such

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 aquacul-

ture practice have received much

attention. The research of prebiotics

in fish nutrition is increasing with the

demand by consumers for safe prod-

ucts produced according the sustain-able practices (Denev et al, 2009).

Aquaculture AndProtection Mechanism

The immune system of the fish

is the primary defence mechanism against infectious disease. A local

immune response in the intestinal mucosa is generated in different steps.

At first macrophages and other phagocytic cells, like dendritic cells, take

up the pathogen (nonspecific immunity), and destroy them with enzymes

and O-radicals, and process it for presentation to local T cells. Phagocytic

Table 1. Total count of bacteria, immunity items and proximate composition of the common carp fingerlingsfed diets containing different levels of ImmunoWall® for 8 weeks.

0.0 g/kg 0.5 g/kg 1.0 g/kg 1.5 g/kg 2.5 g/kg

Parameters

Total counts of bacteria (Log

cfu/g intestine)

9.18

± 0.23

7.78

± 0.26

7.50

± 0.44

7.30

± 0.18

6.96

± 0.17Immunity items

Albumin (g/l) 11.5± 0.4

10.70± 0.7

11.3± 0.5

11.5± 0.5

12.1± 0.8

Globulin (g/l) 13.09± 0.9

12.7± 0.1

13.6± 0.9

14.6± 0.7

15.6± 0.6

Albumin:Globulin ratio 0.82± 0.08

0.84± 0.12

0.83± 0.09

0.78± 0.07

0.77± 0.08

Leucocyte(x10 mm ) 2.01± 0.04

2.28± 0.01

2.25± 0.02

2.24± 0.03

2.15± 0.02

Total protein (g/l) 25.4± 0.55

23.4± 0.35

24.9± 0.40

26.1± 0.25

27.7± 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

Yeast

cell wall

and the

immuno

stimulation

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 ComponentThey 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 host’s 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 degranula- tion 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 Resistanceto 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 MethodsThe 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 differ-

ent 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 ResultsBoth 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

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in the fish fed a diet containing 2.5 g/kg prebiotic. Haematological

parameters and plasma total protein concentration were also signifi-

cantly 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® supplementa-

tion 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 supplement-

ing 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 thecommon 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 compo-

nents 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. van-

namei 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 brewer’s yeast

showed extracellular superoxide anion production of head kidney

macrophages. This prebiotic supplement also improves the fish resist-

ance to A. hydrophila infection, thereby acting as a potent immunestimulant. 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

ImmunoWall®®at levels of 1–1.5 g/kg was beneficial on immune

system, increasing the performance and the survival of C. carpio

fingerlings.

Full citations and references available on request

20 | INTERNATIONAL AQUAFEED | September-October 2014

FEATURE

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September-October 2014 | INTERNATIONAL AQUAFEED | 21

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