Poultry hydrolysates enhance stress resistance and stress tolerance in Pacific white shrimp

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March | April 2014Poultry hydrolysates enhance stress

resistance and stress tolerance in Pacificwhite shrimp

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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Aquaculture is intensifying in

almost all regions of the world

in order to supply the increasing

demands for fish and shrimp.

In Thailand the average stocking den-sity has quadrupled from approximately 40

shrimps/m2 to almost 160 animals/m2.

However, this intensification is often com-

bined to sub-optimal conditions for fish and

shrimps. Problems associated with higher

disease susceptibility have to be faced.

Some of these stress conditions may

be compensated by aquaculture production

practice and/or by an increase use of pharma-ceuticals and antibiotics which in turn affect

food safety.

A ban of antibiotic growth promoters in

many importing countries has forced the indus-

try to search for environmentally friendly alter-

natives. And a natural solution to combat stress

conditions is the use of hydrolyzed proteins.

Fish protein hydrolysates (FH) are consid-

ered as biological active ingredients due to

their beneficial properties to be antioxidative

or antimicrobial (Klompong et al. 2007).The use of FH in diets has proven to

improve growth and performance in fish

for example, Atlantic salmon (Berge and

Storebakken, 1996),

catfish (Herault et

al. 2012) and tiger

prawns P. monodon

(Anggawati et al.

1990) by enhancing

stress resistance and

immunity. Anggawati

et al. (1990) found that

three percent FH was

enough to enhance

shrimp growth.

The immune

boosting effect of FH

is reported due to its

content of peptides,

free amino acids and nucleotides. Although

these peptides can be found in FH, they are

not restricted to marine proteins, only.

Poultry protein derivatives, especially

hydrolyzed poultry proteins, are (as FH) rich

in bioactive peptides, which are able to com-pensate stress conditions.

The benefits of poultry peptidesThe article describes the beneficial effects

of poultry peptides in respect of enhancing

stress resistance/tolerance and immunity in

Pacific white shrimps (Litopenaeus vannamei)

under controlled conditions.

Under controlled laboratory conditions

hydrolyzed poultry peptides (enzymatically

hydrolyzed liver (eL), enzymatically hydro-lyzed meat & bone-mush (eMB)) have been

tested to see the effect on growth perform-

ance in shrimp. The properties of those

peptides are given in Figure 1, indicating the

small molecular size distribution.

Poultry hydrolysates enhance stress resistanceand stress tolerance in Pacific white shrimpby Orapint Jintasataporn Department of Aquaculture, Faculty of Fisheries, Kasetsart University, Bangkok, Thailand and Franz-PeterRebafka, GePro Gefluegel-Protein Vertriebsgesellschaft mbH & Co. KG, Germany

Table 1: Composition of the experimental diets

Raw material Control eL eMB

Fishmeal, tuna 30 7 7

Soybean 9 34 34

Shrimp meal 15 4 4Squid meal 5 1 1

Wheat flour 31.4 29.2 28.85

Hydrolyzed poultryliver

0 5 0

Hydrolyzed poultrymeat & bone

0 0 5

Soy protein isolate 0 6 6

Tuna fish oil 3 3 3

Soya oil 2 2.75 3.1

Squid liver oil 1 1 1

Met + Lys 0 0.45 0.45Mono-cal 1 4 4

Binder 1.7 1.7 1.7

Vitamin-mineralpremix

0.9 0.9 0.9

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The trial was conducted in June 2012 at

Kasetsart University, Bangkok, Thailand, by the

Department of Aquaculture, Faculty of Fisheries,

in a complete randomised design (CRD) with

three treatments and four replicates.

Shrimps were fed with isonitrogenous

diets (see Table 1), including the control (50

percent marine protein) and the test diets

with either five percent hydrolyzed poultry

liver or with five percent hydrolyzed poultry

meat & bone-mush.

Pacific white shrimps (Litopenaeus van-

namei) of 3.1+-0.1g were stocked at a density

of 70 animals/m2 in a 240 litre glass container

(aquarium) in brackish water of 12ppt. During

an eight week period, feed was given three

times per day to match 2-2.5 percent body

weight. Two hours after feeding, the uncon-

sumed feed was collected to determine total

feed consumption.

Growth performance was evaluated every

two weeks by collecting data on average daily

gain, feed consumption, feed conversion ratioand survival rate.

An acute salinity and chemical stress test

with a high or low level of salinity and toxic

concentration of nitrite (NO2-) and ammonia

(NH3+) was conducted at the end of four-

week feeding period.

Under salinity stress 30 shrimps per treat-

ment were transferred from a tank with 12ppt

water salinity to tanks of 20ppt for one week

March-April 2014 | INTERNATIONAL AQUAFEED| 15

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and then moved to with 0ppt or 40ppt saline

water in separate trial for stress test.

In the chemical stress test, 30 shrimpsper treatment were transferred to a tank

with 20ppt salinity for one week and a toxic

concentration of ammonia of 50+- 2ppm

(pH7-8) and in a separate trial of nitrite of

20+-2ppm, respectively.

Nitrite and ammonia concentrations in the

water were controlled daily to maintain the

toxic concentrations. Shrimps were fed oncea day. Mortality rates were recorded daily

during a 10-day test phase.

Typical immunological parameters, such

as hemolymp protein, total hemocyte cell

count, phenol oxidase activity and percentage

of oxyhemocyanin (live cell hemocyte) were

measured. Mortality were recorded daily dur-

ing a 10-day test phase.

After feeding shrimp with the experimental

feed for four weeks, 30 shrimp from eachtreatment were collected to challenge with

White Spot Syndrome Virus (WSSV) and

in a separate trial with Vibrio harveyi by

subcutaneous injection. Mortality rate was

determined daily for a 10-day test phase.

The study was conducted in completely

randomised design (CRD). All data were ana-

lysed by one-way ANOVA. The significance of

differences between means was tested using

Duncans Multiple Range Test at a 95 percent

level of confidence (p

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The activity of hemocyte cell in terms of

phenol oxidase activity and percentage ofoxyhemo-cyanin (live cell hemocyte), was

highest (P

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The immune responses of shrimp

after challenged with white spot syn-

drome virus (WSSV) showed that thegroup of shrimp fed hydrolysed poultry

products had better immune response

than contro l. The mortal ity rate was 100

percent after 72 hours in all treatments

(see Figure 8).

An important sources ofprotein and energy

According to the result from this study, it canbe concluded that growth of shrimps, immune

responses and mortality rate induced by chemical

stress and disease challenge are related to the

usage and uptake of hydrolysed proteins and

peptides.

Generally, rendered poultry by-products

are important sources of protein and energy.

The enzymatic hydrolysis is widely used

to improve and enhance nutritional and func-

tional properties of feedstuff. During the

enzymatical hydrolyses poultry protein is con-

verted into free amino acids, short peptides

and small molecular proteins.

Based on their good functional properties

and nutritive value, these ingredients are able

to replace or complete fishmeal-based diets

in fish and shrimp feed. (Aguila et al., 2007;

Huong et al., 2012).

Several studies have described the anti-

oxidant activity of protein hydrolysates from

chicken (Wu et al., 2005; Rosa et al., 2008),

tuna liver (Je et al., 2009), sardinelle by-prod-

ucts (Bougatef et al., 2010), backbone of Baltic

cod (Zelechowska et al., 2010) and marine

skin gelatins (Alemn et al., 2011).

Furthermore, free amino acid and small

molecular weight compounds released during

the hydrolysis might also act as feed attract-ants for shrimps, increasing feed intake, weight

gain and enhance shrimp immunity (Hardy,

1991).

When shrimp are exposed to high ammo-

nia and nitrite concentration in water, the

immune response is depressed and mortality

is increased.

Hence, robust and healthy shrimp show

a higher immunity resistance by expressing

typical immune parameters, such as hemo-

lymp protein, total hemocyte cell count,

phenol oxidase activity and percentage of

oxyhemocyanin (live cell hemocyte) than

weaker ones.

Althought L. vannameican adapt to a wide

range of salinity, shrimps are more susceptible

to ammonia toxicity. Shrimps under low salin-

ity spend more energy to compensate their

additional energy demand for regulation of

the osmotic pressure (osmoregulation)(Liu

Chun-Hung et al., 2004; Li et al., 2007).

The results are in accordance with previ-

ous studies by Kvale et al. (2002) with pre-

digested protein in Atlantic halibut and Cahu

et al. (1999) describing effects of hydrolysed

protein in sea bass, revealing that survival in

fish can be improved by supplementing (fish)

protein hydrolysates in the diet.

In conclusion, poultry hydrolysates are

able to enhance stress resistance/tolerance in

Pacific white shrimp (Litopenaeus vannamei)

under salinity change, chemical stress and

disease challenge.

MOREINFORMATION:

GePro Gefluegel-ProteinVertriebsgesellschaft mbH & Co. KG

Im Moore 1

49356 Diepholz

Germany

Website: www.ge-pro.de

Email: [email protected]

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