Bol. Inst. Pesca, São Paulo, 44(vol. esp.): 1 - 10, 2017
Doi: 10.20950/1678-2305.2017.1.10
ROUTINE EXPOSURE TO BIOMETRIC PROCEDURES IN FISH FARMING REVEALS
DIFFERENCES IN STRESS RESPONSE IN TAMBAQUI AND HYBRID TAMBATINGA*
Thayssa Cristina Hortences de MORAES
1
; Celma Maria FERREIRA
2
; Kamyla Fernanda
da Silva GAMA
2
; Márcio Aquio HOSHIBA
2
; Jayme Aparecido POVH
3
; Janessa Sampaio
de ABREU
2
ABSTRACT
The physiological stress responses of tambaqui and hybrid tambatinga were evaluated after
subjecting the fish to routine practices in a breeding system such as periodic biometric procedures.
For 270 days of culture, the fish underwent monthly biometric measurements, and at the end of the
period, blood was collected at six sampling times (before, immediately after and 2, 24, 48 and 72 h
after biometric measurements) for the evaluation of physiological indicators of stress. Tambatinga
are more susceptible to stress because they presented higher levels of cortisol and glucose in the
bloodstream after handling and took longer to recover their basal physiological state for these
parameters. However, the low cortisol levels observed in both species suggest that the fish were
familiar with biometric procedures, resulting in a less intense response. Handling led to an increase
in the cellular volume of erythrocytes in tambaqui, resulting in a change in hematocrit and a
decrease in hemoglobin concentration. Hypochloremia was found in both species only 72 h after
handling. Biometric procedures promote hormonal, hematological and hydroelectrolytic changes in
the tambaqui and hybrid tambatinga, but when routinely adopted, at regular intervals, they elicit
stress responses of lower magnitude.
Keywords: Colossoma sp.; cortisol; handling; hematology; fish; Piaractus sp.
EXPOSIÇÃO ROTINEIRA AOS PROCEDIMENTOS BIOMÉTRICOS NA PISCICULTURA
REVELA DIFERENÇAS NA RESPOSTA AO ESTRESSE EM TAMBAQUI E HÍBRIDO
TAMBATINGA
RESUMO
Foram avaliadas as respostas fisiológicas de estresse de tambaqui e híbrido tambatinga, quando
submetidos a práticas rotineiras em sistema de criação, como a realização periódica de biometrias.
Por 270 dias de cultivo os peixes foram submetidos a biometrias mensais e, ao final do período, o
sangue foi colhido em seis tempos de amostragem (antes; imediatamente após; 2; 24; 48 e 72 h após
a biometria) para avaliação de indicadores fisiológicos de estresse. Tambatinga é mais susceptível
ao estresse, pois apresentou maiores níveis de cortisol e glicose na corrente sanguínea após manejo
e levou mais tempo para recuperar seu estado fisiológico basal para estes parâmetros. Contudo, os
baixos níveis de cortisol observados para ambos sugerem que os peixes estavam familiarizados ao
manejo biométrico, resultando em resposta menos intensa. O manejo provocou aumento no volume
celular dos eritrócitos do tambaqui, resultando em alteração no hematócrito e diminuição da
concentração de hemoglobina. Hipocloremia foi verificada em ambos os peixes apenas 72 h após a
realização do manejo. O manejo de biometria promove alterações hormonais, hematológicas e
hidroeletrolíticas no tambaqui e brido tambatinga, mas, quando adotado de forma rotineira, em
intervalos regulares, provoca respostas de estresse de menor magnitude.
Palavras chave: Colossoma sp.; cortisol; hematologia; manejo; peixes; Piaractus sp.
Original Article/Artigo Científico: Recebido em 11/10/2016 Aprovado em 24/03/2017
1
Universidade Federal de Mato Grosso (UFMT), Programa de Pós-Graduação em Ciência Animal. Av. Fernando Corrêa da
Costa, 2367 Boa Esperança CEP: 78.070-900 Cuiabá MT Brazil. e-mail: thayssach.moraes@gmail.com
(corresponding author)
2
Universidade Federal de Mato Grosso (UFMT), Faculdade de Agronomia e Zootecnia. e-mail: celmazootecnista@hotmail.com;
kamyllagama@hotmail.com; tokudazoo@gmail.com; janessabreu@yahoo.com.br
3
Universidade Federal de Mato Grosso do Sul (UFMS), Faculdade de Medicina Veterinária e Zootecnia. Campo Grande
MT Brazil. e-mail: jayme.peixegen@gmail.com
* Financial support: CNPq (Process 447465/2014-7); CAPES (Bolsa de Mestrado)
2 MORAES et al.
Bol. Inst. Pesca, São Paulo, 44(vol. esp.): 1 - 10, 2017
INTRODUCTION
Fish farming in Brazil is currently practiced
with over 30 species with the most diverse feeding
habits, chiefly from tropical climates. Among the
native cultured species, production of round”
fish (species and hybrids of the genus Colossoma
and Piaractus) accounts for 82%, with the tambaqui
(Colossoma macropomum) representing a large part
of this total (IBGE, 2014).
The success of tambaqui in fish farming is
attributed to the uncomplicated production of
fingerlings and their good ability to adapt to
captive conditions, featuring rapid growth and
resistance to low levels of dissolved oxygen in
water, handling and diseases. These fish easily
accept artificial feed when raised in tanks on fish
farms and are able to digest animal and vegetable
protein. The fattening phase lasts 240 to 300 days,
depending on the water availability, using storage
densities between 1 and 1.5 fish per square meter
(ARAÚJO-LIMA and GOMES, 2010), reaching an
average weight of 2.0 kg in less than one year of
cultivation.
In Mato Grosso State, as in others, farmers
have sought to increase the productivity of
tambaqui, but many still choose to produce round
fish hybrids, e.g. the tambatinga, a result of the
cross between female tambaqui (C. macropomum)
and male pirapitinga (Piaractus brachypomus). This
hybrid has gill rakers more developed than the
pirapitinga, which provides greater efficiency in
plankton filtration. This species also easily reaches
commercial weight in a short period and with low
dietary levels of crude protein, which represents
an economy with feeding (SILVA-ACUÑA and
GUEVARA, 2002). In addition, in some regions of
Brazil, many fish farmers prefer to farm hybrid
tambatinga because of attractive body aspects
such as silver color and reddish operculum, which
suits the taste and preference of the consumers,
according some producers. Together with the
tambacu hybrid (female C. macropomum × male P.
mesopotamicus), this is the third most largely
produced fish in the country (IBGE, 2014).
In fish farming, biometric measurements are
taken to evaluate information related to animal
performance such as weight and health status.
Despite being a routine practice under culture
conditions, the procedures may cause stress to
fish, resulting in alterations in their homeostasis
that predispose to the appearance of diseases and
may lead to mortality (URBINATI et al., 2014).
Many fish species respond to stress by
elevating their circulating levels of catecholamines
and corticosteroids (BARTON, 2002). These
primary effects provoke secondary responses
related to energy requirements, including
increased blood glucose and altered electrolyte
homeostasis in blood and tissues. Cortisol and
blood glucose are considered good indicators to
evaluate primary and secondary stress response,
respectively (WENDELAAR BONGA, 1997).
In this regard, blood tests (hematological and
metabolic) can be used to physiologically
characterize a species in its culture environment
and subsequently contribute to studies involving
management of cultured fish. The knowledge of
the stress responses is an important tool to
formulate good management practices so that
handling does not compromise the fish
development in farming. Therefore, in the present
study we analyzed the physiological stress
responses of tambaqui and hybrid tambatinga
subjected to routine practices in fish farming such
as periodic biometric procedures.
MATERIAL AND METHODS
The study was conducted in the Fish Farming
Station of the Experimental Farm at the Faculty
of Agronomy and Animal Science at the Federal
University of Mato Grosso (UFMT). All
experimental protocols were approved by CEUA
(Committee of Ethics in Animal Use/UFMT, case
nº. 23108.069114/2014-85).
The fish were acquired from a commercial
fish farm and transported to the Fish Farming
Station of UFMT and kept in 1-m
3
net cages
identified for tambaqui and tambatinga separately,
until reaching a standard length of 15 cm. On
this occasion, 100 tambaqui (C. macropomum) and
100 tambatinga (female C. macropumum × male
P. mesopotamicus) with an average initial weight
standard error) of 343.06 ± 5.46 g were
individualized with a microchip and distributed
in an 800-m
2
excavated pond with 1.80 m depth
at the stocking density of 0.25 fish per m
2
, with
partial water renewal (average 10%) without
supplemental aeration.
Routine exposure to biometric procedures in fish farming 3
Bol. Inst. Pesca, São Paulo, 44(vol. esp.): 1 - 10, 2017
During the entire culture period, the fish were
fed twice daily with extruded feed (VB
Alimentos
®
) for omnivorous fish containing 32%
crude protein (CP), at a feeding rate varying from
3% to 1% of the live weight, which was adjusted
according to the development stage in which the
fish were at the time.
Biometric measurements were taken monthly
over the course of the experiment (270 days). The
management for the biometric measurements
consisted of first capturing all fish with a trawl
and anesthetizing them with 50 mg L
1
Eugenol
®
(previously diluted in ethanol at 1:4, according to
INOUE and MORAES, 2007) for approximately
three minutes, which was the time necessary for
the fish to show apparent signs of sedation such
as reduced swimming motion, partial loss of
equilibrium and reduced gill ventilation (WOODY
et al., 2002). Next, the fish were removed from the
box with anesthetic and the microchip was
identified to subsequently perform the actual
biometric procedures, which consisted of
determining their weight (Marte Scale - Model AS
2000C) and obtaining morphometric measurements
such as total and standard lengths, head size,
height and width of body. The total and standard
lengths were measured using a fish meter board.
For the other measures, a gauge caliper was used.
After this biometric process, the fish were placed
in a box with clean water and oxygenation until
recovery and were then returned to the pond.
At the end of 270 days of culture, during the
last biometric measurements of the experiment, all
fish were subjected to the same procedures from
the previous months (capture, accommodation in
net cages and anesthesia), but one group of tambaqui
(n = 4) and tambatinga (n = 4) not subjected to any
disturbance was captured before the biometric
measurements and anesthetized and had their
blood collected to serve as control for the
physiological indicators (baseline). Immediately
after the biometric measurements (0 h), the
tambaqui (n = 6) and tambatinga (n = 6) had their
blood harvested by caudal puncture.
Subsequently, the next fish that underwent
biometric measurements, in a random sequence,
were packed in net cages for recovery and
allocated to the culture pond. All net cages were
previously identified with the collection time of
the blood sample after biometric measurements,
and tambaqui and tambatinga samples were
allocated, in the same amount, to the same net
cage, to be collected at the same recovery time, at
random. Next, samples of tambaqui (n = 12) and
tambatinga (n = 12), which had their blood drawn
at 2 h, 24 h, 48 h and 72 h after biometric procedures,
were collected. After blood sample collection, the
fish were returned to the culture pond.
In the blood samples, were evaluated
hematocrit (Microcentrifuge Spin100; centrifugation
for 10 min at 3,300 rpm x g and subsequent
reading using a proper chart); total erythrocyte
count (performed in a Neubauer chamber,
following OLIVEIRA-JUNIOR et al. (2008), with
modifications); and hemoglobin concentration
(hemoglobin cyanide method, using a Labtest
®
commercial kit), analyzed in a Labmax Flex
®
machine. Blood samples were centrifuged (HT
CM-610, 10 min at 3,000 rpm) to separate the
serum for analyses of total protein (biuret method,
using a Labtest
®
commercial kit) and chloride
(using a Labtest
®
commercial kit) and to separate
the plasma for analysis of glucose (enzymatic
methodology by oxidase glucose and Trinder
reagent using a Labtest
®
commercial kit),
performed in a Labmax Flex
®
machine. The serum
was also used for the analysis of cortisol (InVitro
Diagnóstica
®
commercial kit for ELISA
methodology).
During the entire experimental period,
dissolved oxygen and temperature (Digital
Oximeter YSI 55), pH (digital pocket pH meter
Quimis
®
- Q400BD) and un-ionized ammonia
(calculated according to EMERSON et al., 1975)
were monitored weekly, while alkalinity (using
methyl orange indicator solution) and
transparency (Secchi disk) were evaluated
monthly.
A completely randomized design was
employed and data were analyzed by a two-way
analysis of variance (ANOVA), with species
(tambaqui and tambatinga) and sampling times
(baseline, 0 h and 2, 24, 48, and 72 h after
biometric measurements) as the factors. When F
values indicated significance (p<0.05), means were
compared by Tukey's test (TUKEY, 1953). Results
are presented as means ± standard deviation and
data were analyzed by the SAS software.
4 MORAES et al.
Bol. Inst. Pesca, São Paulo, 44(vol. esp.): 1 - 10, 2017
RESULTS
The analyzed water physicochemical
variables were within the acceptable limits for
fish farming, according to MORO et al. (2013)
(Table 1). At the end of the experiment (270
days), the final mean weights were 1,742.40 ±
299.81 g and 1,515.29 ± 333.38 g for tambaqui
and tambatinga, respectively.
The hybrid tambatinga showed a significant
increase in cortisol level soon after the biometric
measurements (0 h), differing statistically from
the tambaqui, and returning to baseline 2 h after
this measurement. However, cortisol level in
hybrid tambatinga rose again 48 h after the end
of measurements and remained significantly
high until the end of the experiment. For the
tambaqui, the lowest level of cortisol was
observed two hours after biometric measurements,
and the highest level was recorded 72 h later, with
no significant difference in relation to baseline
(Figure 1).
Table 1. Water quality parameters (mean ±
standard deviation) during the semi-intensive
culture of tambaqui and hybrid tambatinga.
Parameter
Value
Dissolved oxygen (mg L
-1
)
6.88 ± 1.59
Temperature (°C)
28.19 ± 2.13
pH
7.95 ± 0.81
Transparency (cm)
51.26 ± 15.85
Un-ionized ammonia (NH
3
;
mg L
-1
)
0.02 ± 0.03
Alkalinity (mg CaCO
3
L
1
)
33.18 ± 14.19
Figure 1. Cortisol levels (ηg mL
1
) in tambaqui and hybrid tambatinga subjected to biometric procedures in
semi-intensive fish farming. Baseline: before biometric measurements; 0 h: immediately after biometric
measurements; and 2, 24, 48 and 72 h after biometric measurements. Different letters indicate significant
differences between sampling times; * indicates differences between species (Tukey’s test; p<0.05).
Blood glucose increased in both tambaqui
and hybrid tambatinga soon after the biometric
measurements (0 h) (Figure 2), returning to
homeostasis 2 h after the measurements in the
tambaqui (148.1 ± 8.1 mg dL
1
) and later (in 24 h)
in the tambatinga (97.3 ± 4.9 mg dL
-1
). The blood
glucose levels of the tambaqui were statistically
lower than those observed for the hybrid
tambatinga from 2 to 48 h after the biometric
measurements (Figure 2).
In this study, an increase was found in the
serum concentration of ion chloride 2 h after the
biometric procedures in both tambaqui (199.8 ±
3.8 mEq L
-1
) and hybrid tambatinga (193.9 ± 2.1
mEq L
-1
). The level of this ion was significantly
higher in tambatinga at 24 and 48 h after biometric
procedures in relation to tambaqui, but the
characteristic hypochloremia in response to stress
was found only 72 h after the biometric
measurements in both species (Figure 3).
0
10
20
30
40
50
60
Baseline 0 h 2 h 24 h 48 h 72 h
Cortisol (ɳg mL
1
)
Tambaqui Tambatinga
BC
C
ABC
AB
BC
A
A
AB
AB
C
AB
C
*
*
*
Routine exposure to biometric procedures in fish farming 5
Bol. Inst. Pesca, São Paulo, 44(vol. esp.): 1 - 10, 2017
Figure 2. Glucose levels (mg dL
-1
) in tambaqui and hybrid tambatinga subjected to biometric procedures in
semi-intensive fish farming. Baseline: before biometric measurements; 0 h: immediately after biometric
measurements; and 2, 24, 48 and 72 h after biometric measurements. Different letters indicate significant
differences between sampling times; * indicates differences between species (Tukey’s test; p<0.05).
Figure 3. Chloride levels (mEq L
-1
) in tambaqui and hybrid tambatinga subjected to biometric procedures in
semi-intensive fish farming. Baseline: before biometric measurements; 0 h: immediately after biometric
measurements; and 2, 24, 48 and 72 h after biometric measurements. Different letters indicate significant
differences between sampling times; * indicates differences between species (Tukey’s test; p<0.05).
The total protein concentration increased
significantly soon after the biometric
measurements (0 h) in the hybrid tambatinga
(5.3 ± 0.1 g dL
-1
), differing statistically from the
tambaqui, in which total protein increased 2 h
after the biometric measurements (6.2 ± 0.2 g dL
-1
),
returning to baseline in both species 24 h after the
measurement procedures (Figure 4).
Results for hematological parameters evaluated
here are presented in Table 2. The hybrid tambatinga
did not show significant statistical differences for
hematocrit but differed from the tambaqui, which
showed significantly higher hematocrit values 2 h
after biometric measurements (50.7 ± 2.1%), returning
to baseline 24 h after this measurement procedure
(38.3 ± 7.6%). The hemoglobin concentration
decreased significantly at 2 and 24 h after the
biometric measurements in tambaqui and
tambatinga, respectively, returning to baseline
48 h after the measurement. However, in both
50
100
150
200
250
300
350
400
Baseline 0 h 2 h 24 h 48 h 72 h
Blood glucose (mg dL
1
)
Tambaqui Tambatinga
C
BC
B
C
C
C
C
C
C
A
A
B
*
*
*
0
50
100
150
200
250
Baseline 0 h 2 h 24 h 48 h 72 h
Chloride (mEq L
1
)
Tambaqui Tambatinga
B
B
B
B
B
A
C
C
B
B
B
A
*
*
6 MORAES et al.
Bol. Inst. Pesca, São Paulo, 44(vol. esp.): 1 - 10, 2017
species, a significant decline was found in
hemoglobin concentration 72 h after the biometric
measurements, remaining significantly lower in
tambaqui (7.8 ± 0.2 g dL
-1
) compared with hybrid
tambatinga (9.5 ± 0.6 g dL
-1
). For the tambaqui,
no statistical difference was observed between
sampling times for number of erythrocytes,
although immediately after biometric procedures
(0 h) this parameter was significantly higher (3.7 ±
2.5 cells × 10
6
µL
-1
) in relation to the hybrid
tambatinga (2.7 ± 0.9 cell × 10
6
µL
-1
), which
displayed a higher number of these cells 48 h after
the biometric procedures (5.8 ± 5.5 cells × 10
6
µL
-1
),
returning to homeostasis 72 h after measurements.
Figure 4. Total protein (g dL
-1
) in tambaqui and hybrid tambatinga subjected to biometric procedures in
semi-intensive fish farming. Baseline: before biometric measurements; 0 h: immediately after biometric
measurements; and 2, 24, 48 and 72 h after biometric measurements. Different letters indicate significant
differences between sampling times and * indicates differences between species (Tukey’s test; p<0.05).
Table 2. Hematological parameters of tambaqui and hybrid tambatinga subjected to biometric procedures
in semi-intensive fish farming. Baseline: before biometric measurements; 0 h: immediately after biometric
measurements; and 2, 24, 48 and 72 h after biometric measurements.
Sampling time
Baseline
0 h
2 h
24 h
48 h
72 h
Tambaqui
Ht
38.3 ± 7.6
B
43.8 ± 1.8
AB
50.7 ± 2.1
A
44.8 ± 2.2
AB
41.7 ± 1.2
B
39.6 ± 1.7
B
Hb
12.2 ± 0.4
B
14.6 ± 0.2
A
9.5 ± 0.4
C
3.1 ± 0.3
E
10.5 ± 0.3
BC
7.8 ± 0.2
D
Er
2.5 ± 5.0
A
3.7 ± 2.5
A
2.7 ± 1.6
A
2.9 ± 2.8
A
2.9 ± 4.3
A
2.6 ± 2.2
A
Tambatinga
Ht
46.3 ± 5.5
A
45.0 ± 1.3
A
47.5 ± 2.4
A
42.6 ± 1.6
A
40.8 ± 0.9
A
41.2 ± 1.4
A
Hb
12.3 ± 0.4
AB
13.6 ± 0.4
A
11.0 ± 0.8
ABC
3.1 ± 0.3
D
10.4 ± 0.3
BC
9.5 ± 0.6
C
Er
1.9 ± 0.9
B
2.7 ± 0.9
B
2.9 ± 3.9
B
2.6 ± 1.5
B
5.8 ± 5.5
A
2.8 ± 1.6
B
Means ± standard error. Means followed by common letters in the row do not differ according to Tukey’s test (p<0.05).
Ht: hematocrit (%); Hb: hemoglobin (g dL
1
); Er: erythrocyte count (cells × 10
6
µL
1
).
DISCUSSION
In fish, the primary physiological responses
to different stressors involve neuroendocrine
responses that include the release of
catecholamines from the chromaffin tissue and
stimulus of the hypothalamic-pituitary-interrenal
axis (HPI), culminating in the release of
corticosteroid hormones into circulation
(BARTON, 2002).
The plasma cortisol is the most widely used
indicator of stress in fish, irrespective of their
development stage (WENDELAAR BONGA,
0
1
2
3
4
5
6
7
Baseline 0 h 2 h 24 h 48 h 72 h
Protein (g dL
1
)
Tambaqui Tambatinga
AB
BC
BC
C
A
C
BC
AB
A
C
C
BC
*
*
Routine exposure to biometric procedures in fish farming 7
Bol. Inst. Pesca, São Paulo, 44(vol. esp.): 1 - 10, 2017
1997). Cortisol levels found in this experiment
ranged from 32.89 to 55.35 ɳg mL
-1
for tambaqui
and from 35.99 to 52.10 ɳg mL
-1
for hybrid
tambatinga. Irrespective of the sampling time, the
cortisol values were low as compared with other
studies in which the level of this corticosteroid
was tested in native fish such as tambaqui, pre-
and post-application of acute stressors like capture
(TAVARES-DIAS et al., 2001) and transport
(GOMES et al., 2003a; c; CHAGAS et al., 2012).
Although the biometric practice exposes the
fish to sequential stressors that often cause stress,
in the present study, the biometric procedures
were performed every month during the 270 days
of culture, and the fish thus became accustomed
to these measurements. The continuous cortisol
production by the interrenal cells as a result of
repeated exposure to stressors could negatively
regulate the HPI axis by negative feedback,
reducing neuroendocrine responses (BARTON,
2002).
Rainbow trout (Salmo gairdneri) juveniles
captured daily for 10 weeks displayed a lower
plasma cortisol level at the end of this period,
indicating a possible desensitization of the HPI
axis due to the overall habituation of the fish to
repeated disturbances (BARTON et al., 1987).
Tambacu hybrids (female C. macropomum × male
P. mesopotamicus) also showed decreased cortisol
levels one hour after being subjected to the
application of consecutive capture stimuli
(MARTINS et al., 2002).
Despite the low cortisol values observed in
this study for both species, immediately after the
biometric procedures (0 h), the tambatinga presented
significantly higher cortisol levels than the
tambaqui, suggesting that this primary stress
response may be of greater magnitude in hybrid
as compared with pure species. The physical
strain by the tambatinga during the biometric
procedures might have contributed to a more
intense and later release of cortisol, which, unlike
in tambaqui, rose again 48 h after biometric
procedures. According to HOSHIBA et al. (2009),
during physical exercise, the elevation in plasma
cortisol is delayed. The plasma cortisol increase is
usually not observed before 30-60 min after the
end of exercise (MILLIGAN and WOOD, 1987),
and release peaks occur up to 1-2 h later (GAMPERL
et al., 1994).
Responses secondary to stress include
metabolic, ionic and hematological changes
related to physiological adjustments in the
metabolism, respiration, hydromineral balance,
immune function and cellular responses
(WENDELAAR BONGA, 1997; BARTON, 2002).
Increased plasma glucose levels have been
reported after acute and chronic stress due to
glycogenolysis and gluconeogenic effects caused
by catecholamines and cortisol, respectively
(WENDELAAR BONGA, 1997). In this regard, the
glucose is mobilized as an energy source, and this
was found in the present study. As described for
tambaqui (GOMES et al., 2003a; 2003c) and other
native fish species such as pirarucu (GOMES et al.,
2003b) and matrinxã juvenile (URBINATI et al.,
2004), in this study, there was an increase in blood
glucose immediately after the stimulus to stress
(0 h), but the time for return was different among
the species, with later return to baseline for
tambatinga (24 h) than tambaqui (2 h). In addition,
the blood glucose level of the hybrid was
significantly higher than that of the pure species
up to 48 h after the biometric procedures.
Alterations in circulating glucose caused by a
stressor agent may vary depending on factors like
species, size, sex, line and characteristics of the
stressor agent, e.g. type, severity and intensity. In
the same way, the capacity of return to baseline
conditions may vary when the stressor stimulus is
interrupted (TAKAHASHI et al., 2006). The results
suggests that tambatinga is more susceptible to
stress than tambaqui, since the former presented
higher cortisol and glucose concentration in blood
circulation after handling and took longer to
recover the basal physiological state for these
parameters.
Hydroelectrolytic imbalances have been
observed as a stress response in fish
(McDONALD and MILLIGAN, 1997). Likewise,
alterations in hematocrit and hemoglobin
concentration may evidence hemodilution or
hemoconcentration caused by stressful situations
(MORGAN and IWAMA, 1997; BOSISIO et al.,
2017). In this study, it was evident that these
alterations occurred for both species, especially
two hours after biometric measurements. The
increase in cell permeability during stress
resulted in changes of fluid from the plasma to
the intracellular compartment, concentrating more
8 MORAES et al.
Bol. Inst. Pesca, São Paulo, 44(vol. esp.): 1 - 10, 2017
molecules in the blood plasma, which explains the
significant increase in total protein and plasma
chloride at this sampling time. The influx of water
into the cell contributed to increasing their
volume, explaining the increase in hematocrit and
decrease in hemoglobin concentration, especially
in the tambaqui at this sampling time (2 h). As
documented by MORGAN and IWAMA (1997),
the mobilization of catecholamines into the fish
blood during stress causes cell swelling, resulting in
hemoconcentration in several freshwater species
(McDONALD and MILLIGAN, 1997).
After a stress condition, serum sodium and
chloride levels decline and potassium and calcium
levels are modified. When freshwater fish face
adverse conditions, there is a loss of chloride ions
from their blood to the water as well as excessive
hydration of the body, which causes these fish to
expend additional energy to maintain or
reestablish the osmoregulatory balance (HOSEINI
et al., 2016). In both species investigated in this
study, the change in cell permeability of the gill
epithelium seems to have occurred later, since
the characteristic hypochloremia (loss of chloride
ions to the aquatic environment) in response to
stress was found only 72 h after the biometric
measurements.
These results may vary depending on the
fish species and the stressor applied. In pacu
(P. mesopotamicus), ABREU et al. (2009) found a
significant decrease in chloride levels 60 min after
the fish had been subjected to capture, without
return to homeostasis for up to 24 h. FAGUNDES
and URBINATI (2008), however, did not find
changes in serum chloride levels in Spotted
sorubim (Pseudoplatystoma corruscans) in up to 48 h
after stress from capture and transport. A slight
decreased in plasma total protein concentration
was found for pacu (P. mesopotamicus) after transport
(FEITOSA et al., 2013), and no significantly
difference for this parameter was observed for
matrinxã (Brycon amazonicus) after the challenge of
stress from capture and exposure to air (ABREU
and URBINATI, 2006).
The hematological responses shown by the
tambaqui observed in the current study indicate
that the biometric procedures did not lead to an
increase in the number of red cells, but rather in
the cellular volume of erythrocytes, resulting in
alterations in hematocrit. The decreased hemoglobin
concentration as well as the increased
concentration of total serum protein observed at
these sampling times in tambaqui reinforce a
change of fluid from the plasma to the
intracellular compartment of the erythrocytes as a
result of the increased permeability of the cell
membrane brought about by the release of
catecholamines during stress (MELO et al., 2009).
For the tambatinga, these hematological alterations
were not as significant as in the tambaqui. The
increase in number of red blood cells of the
tambatinga at 48 h was an isolated alteration
resulting from the release of erythrocytes to the
blood stream by spleen contraction, possibly
caused by the increase in catecholamine
concentration during stress.
CONCLUSIONS
Tambatinga is more susceptible to stress than
tambaqui, since the former presented higher
cortisol and glucose concentrations in the blood
circulation after the management and took longer
to recover the basal physiological state for these
parameters. Biometric procedures cause
physiology changes in tambaqui and hybrid
tambatinga, but when adopted routinely, at
regular intervals, they lead to stress responses of
lower magnitudes, with levels below those found
in the literature.
ACKNOWLEDGMENTS
The authors thank UFMT for providing the
experimental site, equipment, materials and
reagent used; the VB Alimentos
®
company for
donating the extruded feed used in the
experiment; the Lufada fish industry for donating
the fingerlings; and CNPq/CAPES for the
financial support.
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