Bol. Inst. Pesca, São Paulo, 44(vol. esp.): 85 - 92, 2017
Doi: 10.20950/1678-2305.2017.85.92
FISH PROTEIN HYDROLYSATE AS AN INGREDIENT IN DIETS FOR Arapaima gigas
JUVENILES*
Melquisedeque da Silva RIBEIRO
1
; Flávio Augusto Leão da FONSECA
1,
2
; Marieta
Nascimento de QUEIROZ
1
; Elizabeth Gusmão AFFONSO
1,
3
; Luís Eugénio Castanheira da
CONCEIÇÃO
4
; Ligia Uribe GONÇALVES
1,3
ABSTRACT
This study evaluated the dietary inclusion of a fish protein hydrolysate (FPH) derived from from
tilapia trimmings, on physiological and growth parameters of juveniles of Arapaima gigas. A total of
180 arapaima juveniles (91.4 ± 2.7 g) were used in a complete randomized design with six
treatments (n = 3). Fish were fed to apparent satiation four times a day for eight weeks, with diets
containing increasing inclusion levels of FPH (0, 4, 8, 12, 16 and 20%). FPH diets did not affect
growth and hemato-biochemical parameters of arapaima juveniles. The FPH from tilapia
trimmings seems to be a suitable ingredient for arapaima over 90 g feeds, at least up to 20%
inclusion level. No bioactive effects of the FPH could be detected.
Keywords: alternative ingredient; tilapia trimmings; carnivorous fish
HIDROLISADO PROTÉICO DE PEIXE EM DIETAS PARA JUVENIS DE Arapaima gigas
RESUMO
Este estudo avaliou a inclusão na dieta de um hidrolisado de proteína de peixe (FPH) derivado de
aparas de tilápia sobre parâmetros fisiológicos, crescimento e composição proximal do músculo de
juvenis de Arapaima gigas. Cento e oitenta juvenis de arapaima (91,4 ± 2,7 g) foram utilizados em
delineamento inteiramente casualizado, com seis tratamentos (n = 3). Os peixes foram alimentados
até saciedade aparente quatro vezes por dia durante oito semanas, com dietas contendo níveis
crescentes de inclusão de FPH (0, 4, 8, 12, 16 e 20%). As dietas de FPH não afetaram o crescimento e
os parâmetros hemato-bioquímicos de juvenis de arapaima. O FPH de aparas de tilápia parece ser
um ingrediente adequado para rações de arapaima acima de 90 g, pelo menos até 20% de inclusão.
Nenhum efeito bioativo da FPH pode ser detectado.
Palavras-chave: ingrediente alternativo; aparas de tilápia; peixe carnívoro
Original Article/Artigo Científico: Recebido em 15/11/2016 Aprovado em 05/09/2017
1
Universidade Nilton Lins, Programa de Pós-Graduação em Aquicultura. Parque das Laranjeiras - Av. Prof. Nilton Lins,
3259 CEP: 69058-580 Flores Manaus AM Brazil. e-mail: melquisedequesr@gmail.com; marie_quei@hotmail.com
2
Instituto Federal de Educação, Ciência e Tecnologia do Amazonas (IFAM) - Campus Zona Leste. Av. Cosme Ferreira, 8045
CEP: 69086-475 Gilberto Mestrinho Manaus AM Brazil. e-mail: flaviofonseca@ifam.edu.br
3
Instituto Nacional de Pesquisas da Amazônia (INPA), Coordenação de Tecnologia e Inovação. Av. André Araújo, 2.936
CEP: 69.067-375 Petrópolis Manaus AM Brazil. e-mail: pgusmao@inpa.gov.br; ligia.goncalves@inpa.gov.br
(corresponding author)
4
SPAROS Lda. Área Empresarial de Marim Olhão Portugal. e-mal: luisconceicao@sparos.pt
* Financial support: Fundação de Amparo à Pesquisa do Estado Amazonas (FAPEAM) (Process n° 062.00676/2015)
86 RIBEIRO et al.
Bol. Inst. Pesca, São Paulo, 44(vol. esp.): 85 - 92, 2017
INTRODUCTION
Arapaima gigas is one of the most important
native species for Brazilian aquaculture. It is a
carnivorous species with big growth rates, high
fillet yield and high market value, which makes it
very attractive for fish farming (IMBIRIBA, 2001;
OLIVEIRA et al., 2012; VALLADÃO et al., 2016).
However, there are limitations in juveniles supply
mainly due to absence of techniques for artificial
reproduction and high mortality rates observed in
arapaima early life (NÚÑEZ et al., 2011; LIMA et al.,
2015). Diets with an optimal cost-effectiveness and
reducing susceptability to oportunistic pathogens
are also lacking for this species. The use of diets
with high biological value and immunostimulant
action ingredients could reduce growth losses and
mortality rates due to parasites.
Fish protein hydrolysates (FPH) have been
described to possess bioactive peptides and amino
acids which can act as antibiotics, antibacterial
agents, antioxidants, and regulators of the activity
of certain digestive enzymes (GILL et al., 1996).
FPH are considered potential ingredients for the
aquafeed industry, due to its high protein content,
as well as having flavoring action (CONCEIÇÃO
et al., 2012; HE et al., 2013). The inclusion up to
10% FPH in juvenile diets for croaker, Pseudosciaena
crocea, provided improvements in growth and
immunological parameters (TANG et al., 2008).
However, these benefits are not present for all
species. The addition of 15% and 25% FPH in diets
for juvenile turbot, Scophthalmus maximus, reduced
protein and energy digestibility due to
gastrointestinal disorders (OLIVA-TELES et al., 1999).
The objective was to evaluate the dietary
inclusion level of protein fish hydrolyzate on
growth and haematological parameters of arapaima
juveniles.
MATERIAL AND METHODS
This study has been approved by the Ethical
Committee of Animal Experimentation and
Research of the Instituto Nacional de Pesquisas da
Amazônia (INPA), Manaus, Amazonas, Brazil
(Protocol Number 016/2016).
Arapaima juveniles (91.4 ± 2.7 g; 23.0 ± 0.6 cm)
were simultaneously housed (10/0.15 m
3
) in 150 L
tanks, adding up 18 experimental units with
open water system and constant aeration, under
a full randomised design with six treatments
and three replicates, at Fish Farming Station of
Instituto Nacional de Pesquisas da Amazônia
(INPA, Manaus, Brazil).
Temperature (29.09 ± 0.64 ºC), dissolved
oxygen (6.49 ± 0.51 mg L
-1
) (YSI Pro20 Dissolved
Oxygen Meter) and pH (5.84 ± 0.43) (YSI Pro10
pH) of water were daily monitored. Every week
water total ammonia (0.00 mg L
-1
) and nitrite
(0.00 mg L
-1
) concentrations were measured
according to VERDOUW et al. (1978) and BOYD
and TUCKER (1992) respectively. Water parameters
remained within the comfort range for arapaima
(CAVERO et al., 2003; NÚÑEZ et al., 2011;
OLIVEIRA et al., 2012).
Six isonitrogenous (46% CP) and isocaloric
(4200 kcal kg
-1
) diets were formulated with
increasing levels (0, 4, 8, 12, 16 and 20%) of fish
protein hydrolysate from tilapia trimmings (FPH)
(Table 1). Fish protein hydrolyzed, ingredients
and feedstuff were analyzed to centesimal
composition (AOAC, 2005) at INPA’s Fish Nutrition
Laboratory.
The ingredients were milled, homogenized
(0.9 mm), hydrated (20% water: volume), and
pelleted (4-5 mm). The diets were dried in forced
air ovens (50 ° C, 24 h) and stored (-4 °C) until use.
Fish were fed experimental diet four times a
day (8:00, 11:00, 14:00 and 17:00 h) during eight
weeks. At the end of growth assay, fish were
fasted during 24 h.
The following performance parameters were
evaluated according to the formulas (NRC, 2011):
Survival Rate (SR) = (final number of fish x
100)/initial number of fish;
Individual Weight Gain (IWG) = final weight (g)
initial weight (g);
Feed Intake (FI) = (initial feed weight (g) - final
feed weight (g))/number of fish;
Feed Conversion Ratio (FCR) = feed intake
(g)/weight gain (g);
Daily Weight Growth (DWG) = WG (g)/days of
experiment (days);
Relative Growth Rate (RGR) = (e
g
-1) x 100;
where: e = nepper number; g = (ln(final weight)
ln(inicial weight))/(length of the assay period);
Hepatosomatic Index (HSI) = liver weight
(g)/body weight (g) x 100.
Fish protein hydrolysate as an ingredient in diets for Arapaima gigas 87
Bol. Inst. Pesca, São Paulo, 44(vol. esp.): 85 - 92, 2017
Table 1. Percentage and chemical composition of experimental diets for juvenile Arapaima gigas fed diets
containing increasing levels of fish protein hydrolysate.
a
FPH amino acid composition (g 100 g
1
): arginine (2.8); histidine (0.65); isoleucine (1.54); leucine (2.87); lysine (3.17);
methionine (1.01); phenylalanine (1.66); threonine (1.64); tryptophan (0.31); valine (1.99); cysteine (0.3); tyrosine (1.12);
glutamic acid (5.42); glycine (4.35); serine (1.53); proline (2.66); alanine (3.02); aspartate (3.61); taurine (0.32);
hidroxiprolina (1.28);
b
Chemical composition: dry matter (92%); ash (8.6%); lipids (1%); protein (41.25%);
c
Premix
Nutron®. Brazil: Mn 26 mg; Zn 140 mg; Fe 100 mg; Cu 14 mg; Co 0.2 mg; I 0.6 mg; Se 0.6 mg. A 10000 UI; D3 4000 UI;
E 100 mg; K 5 mg; B1 25 mg; B2 25 mg; B6 25 mg; B12 30 mg; niacin 100 mg; folic acid 5 mg; panthotenic acid 50 mg;
biotin 0,8 mg; colin 2000 mg; inositol 50 mg; C 350 mg.
Blood samples (1.5 mL) were taken by caudal
venipuncture (6 fish tank
-1
), with ethylenediamine
tetraacetic acid (EDTA-10%) coated syringes for
hematological and biochemical parameters
evaluation. From these a subsample (3 fish tank
-1
)
were euthanized with anesthetic overdose (0.4 mL
eugenol L
-1
water) to collect the liver and calculate
the HSI. From these subsamples, fish muscle
were collected and frozen to posterior chemical
composition analysis.
The blood samples were used for hematocrit
(GOLDENFARB et al., 1971) and hemoglobin [Hb]
(BLAXHALL and DAISLEY, 1973) percentage
analysis. Erythrocyte number (RBC) was counted
in a Neubauer chamber in diluted blood (1:200)
(NATT and HERRICK, 1952). Hematimetric
parameters such as mean corpuscular hemoglobin
concentration (MCHC), mean corpuscular
hemoglobin (MCH) and mean corpuscular volume
(MCV) were obtained according to WINTROBE
(1934). Determination of glucose, total protein,
cholesterol, triglycerides and albumin from plasma
were performed after whole blood centrifugation
( C, 12000 rpm/180 s), using commercial kits and
spectrophotometric readings.
Data with parametric distribution (FI, FCR,
DWG, RGR, SR, Hb, Ht, MCHC, glucose,
protein, cholesterol, triglycerides and albumin)
were analysed by one-way ANOVA (p<0.05).
Data with nonparametric distribution