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FEATURE

Nutraceutics for the control of
bacterial coldwater disease
by C. Fernández-Álvarez, Facultad de Biología (CIBUS), Y. Torres-Corral, Facultad de Biología (CIBUS), G. Coscelli, Facultad de
Ciencias Veterinarias, Universidad Nacional de Rosario, Argentina, A. R. Sánchez-Arévalo, Lípidos Toledo S.A; Madrid, Spain,
A. Martínez, Lípidos Toledo S.A; Madrid, Spain, M.I. Quiroga, Facultad de Veterinaria, Universidad de Santiago de Compostela,
Spain, Y. Santos, Facultad de Biología (CIBUS)

T

he aquaculture industry has
developed significantly over recent
decades and is, today, one of the
fastest-growing food production
sectors in the world. One of the most
important problems that affect aquaculture
is the appearance of infectious diseases.
Among bacterial diseases affecting cultured
salmonid fish, Bacterial Cold-Water Disease,
caused by Flavobacterium psychrophilum,
produces high mortality and morbidity and
consequently, economical losses worldwide.
Attempts to control
this situation through
the use of antimicrobials
have been limited due to
the rapid appearance of
antibiotic-resistant bacteria.
Moreover, the determinants of antibiotic resistance that have emerged
and selected in this aquatic
environment have the
potential of being transmitted by horizontal gene
transfer to bacteria of the
terrestrial environment,
including human and animal
pathogens.
Evidence indicating that
antibiotic-resistant bacteria
and antibiotic resistance
determinants pass from the

aquatic to the terrestrial environment has
resulted in a drastic restriction of the use of
antibiotics in aquaculture in many countries. For
this reason, it is important to find new products
with antimicrobial activity without the aforementioned undesirable effects of antibiotics.
In this sense, natural food additives or phytobiotics, which combined different mechanisms of action against pathogenic bacterial
species (bactericidal/ bacteriostatic activities,
Quorum Sensing inhibition), are potential
candidates for the development of prevention
strategies in aquaculture. This work describes

the effect of the dietary administration to
rainbow trout of the phytobiotic Liptofry
(Liptosa S.A, Spain) in the improvement
of disease resistance against Flavobacterium
psychrophilum.
In this study 4000 healthy rainbow trout,
distributed in two experimental groups (2000
individuals/each) were maintained in an aquaculture facility of the north of Spain, until
its transfer to the aquarium of the Faculty
of Biology of the University of Santiago de
Compostela. Rate of food supply of both
groups was established based on the requirements of the fish, taking as
a reference the parameters
of size and weight of the
specimens and the temperature of the water.
• Group 1: Fish fed with
feed supplemented with
the phytobiotic Liptofry
manufactured by Lípidos
Toledo SA (Liptosa SA,
Spain).
• Group 2: Fish fed with
no supplemented feed
(control fish).
The usefulness of
feeding the fish with diet
supplemented with phytobiotics in the prevention
of Bacterial cold-water disease (BCWD) in rainbow
trout was determined by

14 | INTERNATIONAL AQUAFEED | November-December 2015

FEATURE

of both experimental groups showed similar
lesions, characterised by a moderate inflammatory response at the coelomic cavity,
necrotic areas affecting the spleen, liver, and
pancreatic lobes, and degenerative lesions of
renal tubules, showing pyknosis and dequamation of epitelial cells (Figure 2),
Results of this study, suggest that the use
of
feed
supplemented with the phytobiotic
thickness,
and
stained
with
hematoxylin-eosin
experimental infection, using a virulent strain
RBT4.1.04 of the species F. psychrophilum. (H-E), Phosphotungstic acid-haematoxylin Liptofry in the diet of rainbow trout may conWith this aim, 120 fish of each experimental stain (PTAH) and Periodic acid–Schiff stain tribute to the prevention of diseases caused
by the bacterial pathogen Flavobacterium
group (group 1 and group 2) were anaesthe- (PAS) for light microscopy observation.
In our study, fish fed with the Liptofry sup- psychrophilum. Thus, these diet supplements
tised by immersion in tricaine methane sulfonate (MS-222, Sigma) (60mg/L) and infected plemented diet (group 1) showed an average could be used as an alternative to antibiotics
by intra-peritoneal injection of a bacterial sus- cumulative mortality of 27 following challenge or vaccines in the control and/or prevention of
pension containing 109 CFU/mL. Differences experiment, compared to 62 percent mortal- infectious diseases. Although distribution of the
in survival between fish fed with phytobiotics ity observed in control fish (group 2) (Table inflammatory response in fish of groups 1 and
(Group 1) and control fish (Group 2) after 1). The survival of fish fed with the phytobiotic 2 infected with Flavobacterium psychrophilum
challenge with Flavobacterium psychrophilum Liptofry was significantly higher than those of was similar, the severity and extent of lesions,
and the number of animals affected were
were analysed by Chi-square test (P<0.05) the control group (p<0.05).
Histopathological analysis showed that fish higher in the control group. These findings
(Figure 1).
might correlate with significant
Histopathological
analydifferences of mortality in groups
sis was performed in order
Table 1. Experimental infection with F. psychrophilum to evaluate the
one and two.
to evaluate the effect of the
effect of the Liptofry supplemented diet in the prevention of BCWD
administration of the phytoInoculated dose
Nº death fish/
biotic Liptofry in fish tissues.
Acknowledgements
Experimental group
% Mortality
(CFU/fish)

inoculatedAfish| Version: 1
ulture | 2015
Adcontrol
Campaign
| Theme: Ad-3 Barley Dunne | Design:
TissueHatchery
samples from
and
This work was supported by
Liptofry-feeded
fish were taken
Grant CDTI-IDI-20140147 from
F | Size: Half
Page | Dimensions:
190mm X 132mmGroup 1
Fish treated with the
4.2 x 108
32/120
27
at 24, 48, 72 y 96 h after experithe Centro para el Desarrollo
phytobiotic Lyptofry
mental infection. Bouin’s fixed
Tecnológico e Industrial (CDTI),
Group 2
4.2 x 108
74/120
62
samples were embedded in parMinisterio de Economía y
Control fish
affin wax, sectioned at 3 μm in
Competitividad, Spain

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November-December 2015 | INTERNATIONAL AQUAFEED | 15

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