Bull Environ, Contam. Toxicol. (1993) 50:659--664
9 1993 Springer-Verlag New York Inc.
i
Environ.rner~..al
Effect of Detergent on the Response to Temperature and
Growth of Grass Carp, Ctenopharyngodon iclella
Guillermina Alcaraz, Carlos Rosas, and Sonia Espina
Ecophysiology Laboratory, Department of Biology, Faculty of Science, National
Autonomous University of Mexico, M6xico, D.F., CP 04510, Mexico
The increase in urbanization in the last decades has
p r o d u c e d an increase in discharges
of domestic and
industrial residues.
During this period, detergents
have become a severe problem
in aquatic systems. These
substances are rarely found in c o n c e n t r a t i o n s lethal to
aquatic
organisms;
however,
they
cause
stress
and
interfere with their fitness.
Sublethal concentrations of surfactants harm the gill
epithelium of fish by changing the lipid c o m p o s i t i o n of
the tissue and affecting the production of mucus.
They
also partially destroy the epithelium chloride cells
(Schmid and Mann 1961; Abel and Skidmore 1974).
The
deterioration
of
the
gill
epithelium
alters
the
respiratory function and the hydromineral balance in
these organisms (Rosas et al. 1988).
The presence of
detergents in the environment has also been seen to
harm the peripheral nerve receptors of fish which in
turn causes changes in feeding and t h e r m o r e g u l a t o r y
behavior
(Bardach et al. 1965; Olgivie and A n d e r s o n
1965).
Changes
in
the
energy
balance
of
C t e n o p h a r y n q o d o n idella (Val.) subjected to sublethal
c o n c e n t r a t i o n of detergents have been recently reported
(Espina et al. 1986).
These responses together with
selection and avoidance of temperature have survival
value
as they
affect
the
functional
relationships
between
the organism
and its environment,
and are
ecologically
meaningful
as a result
(Sprague
1971;
Prosser and Nelson 1981).
Several authors have observed changes in the
selection
of, as well as in the tolerance to t e m p e r a t u r e on the
part of fish subjected to sublethal c o n c e n t r a t i o n s of
c o n t a m i n a t i n g agents.
However, little is known about
the effect of detergents on fish responses.
The effect
of a detergent of domestic use on the thermic responses
of the juveniles of the herbivorous carp C. idella was
Send reprint requests
above address.
to
Guillermina
659
Alcaraz
at
the
e v a l u a t e d in this work.
This species was s e l e c t e d as a test o r g a n i s m b e c a u s e of
its i m p o r t a n c e in a q u a c u l t u r e in this country.
MATERIALS AND METHODS
J u v e n i l e s of the h e r b i v o r o u s carp C. idella (1.03-2.60
g wet weight) o b t a i n e d from a c o m m e r c i a l h a t c h e r y were
kept in the l a b o r a t o r y in glass a q u a r i a at a d e n s i t y of
ten fish per 30 L, in dechlorinated, a e r e a t e d w a t e r
w i t h an a l k a l i n i t y ~ f 136-139 mg CaCO3/L.
Fish w e r e
m a i n t a i n e d at 24 - l~
for 1 wk after w h i c h the
t e m p e r a t u r e was raised l~
day up to 29 t_ loC.
This t e m p e r a t u r e is d i r e c t l y related to the carp's
thermal p r e f e r e n d u m when m e a s u r e d against a h o r i z o n t a l
g r a d i e n t in the l a b o r a t o r y (Alcaraz 1989).
Fish w e r e
fed (8% of their body w e i g h t per day) g r o u n d carp
p e l l e t s c o m b i n e d with lucern (75:25). P h o t o p e r i o d was
m a i n t a i n e d at 12 h of light:12 h of darkness.
S u b l e t h a l c o n c e n t r a t i o n s of c o m m e r c i a l d e t e r g e n t (41.9%
s o d i u m alkyl aryl sulphonate, AAS) of 0.0 (control),
5.0, 8.0 and 12.6 m g / L were used.
T h i r t y fish were
e x p o s e d (21-25 d) in separate 30 L a e r e a t e d aquaria at
29-C, and two thirds of the w a t e r v o l u m e was c h a n g e d
daily after feeding (Espina et al. 1986).
Behavioral
responses
of
fish
to
temperature
were
e v a l u a t e d along a h o r i z o n t a l thermal g r a d i e n t (Alcaraz
1989).
G r a d i e n t was e s t a b l i s h e d in an a s b e s t o s tube.
Fish were i n t r o d u c e d into the g r a d i e n t (19-38~
sector
at a t e m p e r a t u r e similar to that of a c c l i m a t i o n (29~
and o b s e r v a t i o n s started 40 min later.
The fish
positions
along
the
thermal
gradient
and
the
temperature
were
recorded
for
the
control
and
e x p e r i m e n t a l groups every i0 min for a p p r o x i m a t e l y 3
hr.
Five carps were m e a s u r e d
at a time.
Four
r e p l i c a t i o n s of each t r e a t m e n t w e r e c o n d u c t e d (n=80) at
the same time of day in order to avoid diel f l u c t u a t i o n
effects on fish preferenda.
Superior
incipient
lethal
temperature
(SILT)
was
e s t a b l i s h e d a c c o r d i n g to K i l g o u r et al.
(1985)
to
e v a l u a t e thermal t o l e r a n c e of the j u v e n i l e g r a s s carps,
w h i c h is the same t e m p e r a t u r e at w h i c h 50% of the test
fish died.
Ten fish of all e x p e r i m e n t a l c o n d i t i o n s
w e r e a b r u p t l y t r a n s f e r e d from w a t e r at 29~ to a e r e a t e d
w a t e r baths at seven d i f f e r e n t temperatures.
Fish that
r e m a i n e d alive for more than 120 min w e r e discarded.
R e s i s t a n c e times were r e g i s t e r e d for i n d i v i d u a l fish in
each bath.
Data were used to c a l c u l a t e m o r t i f i c a t i o n
rates a c c o r d i n g to the above m e n t i o n e d authors.
660
G r o w t h rates from fish s u b j e c t e d to d i f f e r e n t d e t e r g e n t
c o n c e n t r a t i o n s (0, 5, 8 and 12.6 mg/L), w e r e c a l c u l a t e d
from the d i f f e r e n c e b e t w e e n the initial and the final
wet w e i g h t a f t e r 21 days.
V a l u e s w e r e e x p r e s s e d as
mg/day.
For the s t a t i s t i c a l analysis, m e d i a n v a l u e s of fish
thermal p r e f e r e n c e and c o n f i d e n c e i n t e r v a l s (95%) w e r e
c a l c u l a t e d u s i n g the S T A T G R A P H I C S s t a t i s t i c a l p r o g r a m
(1985).
This p r o g r a m also f a c i l i t a t e d
c a l c u l a t i o n of
the r e g r e s s i o n line p a r a m e t e r s fitted by the least
squares m e t h o d used in the SILT calculation.
In o r d e r
to i d e n t i f y s i g n i f i c a n t d i f f e r e n c e s the K r u s k a l - W a l l i s
non p a r a m e t r i c test was c a r r i e d out (Zar 1974).
RESULTS AND DISCUSSION
A
survival
of
maintenance period
were
subjected
concentrations.
detergent affected
t e m p e r a t u r e of the
idella.
100%
was
registered
during
the
as well as d u r i n g the time the fish
to
the
different
detergent
However,
exposure
(21 d)
to
the
g r o w t h and c h a n g e d the r e s p o n s e s to
j u v e n i l e s of the h e r b i v o r o u s carp C.
The final t e m p e r a t u r e p r e f e r e n d u m of the u n e x p o s e d fish
m e a s u r e d by the acute m e t h o d was 29~
A t e n d e n c y to
prefer
higher
temperatures
as
the
detergent
c o n c e n t r a t i o n in the e n v i r o n m e n t increased was o b s e r v e d
(Table i), a l t h o u g h only at the g r e a t e s t c o n c e n t r a t i o n
were the d i f f e r e n c e s b e t w e e n the control g r o u p and the
o t h e r e x p e r i m e n t a l groups s i g n i f i c a n t (P < 0.05).
Superior
incipient
lethal t e m p e r a t u r e
(SILT)
of C.
idella
subjected
to
the
detergent
significantly
d e c r e a s e d (P < 0.05) with an increase in the d e t e r g e n t
c o n c e n t r a t i o n in the e n v i r o n m e n t (Table i).
The v a l u e
of SILT was d i f f e r e n t from that of the control at each
c o n c e n t r a t i o n (P < 0.05).
T h e r e w e r e not any d i f f e r e n c e s in the initial fish
w e i g h t of control and e x p e r i m e n t a l g r o u p s (P>0.05). The
g r o w t h rate (mg/day) of the fish in the control
group
was s i g n i f i c a n t l y g r e a t e r (P<0.05) than that of the
carps s u b j e c t e d to the c o n t a m i n a t i n g agent (Table 2).
A m o n g these it was p o s s i b l e to o b s e r v e an i m p o r t a n t
d e c r e a s e in g r o w t h when the d e t e r g e n t c o n c e n t r a t i o n
increased.
For the control group a 31% increase in
w e i g h t was r e g i s t e r e d
in 21 d, in c o n t r a s t a 16%
increase
in
weight
the
carps
subjected
to
a
c o n c e n t r a t i o n of 5 m g / L of detergent, and a 4.7 and
3.7% increase in w e i g h t those s u b j e c t e d to the g r e a t e s t
c o n c e n t r a t i o n s (8 and 12.6 mg/L).
661
T a b l e i. M e d i a n v a l u e s and 95% c o n f i d e n c e
limits of
p r e f e r r e d t e m p e r a t u r e (PT) and s u p e r o r i n c i p i e n t lethal
t e m p e r a t u r e (vC) of C. idella e x p o s e d 21 d to d i f f e r e n t
detergent concentrations
(mg/L). N u m b e r of fishes in
parentheses.
Detergent
PT
0.0
30.00
29.7 - 30.3
(20)
30.97
30.5 - 31.4
(20)
30.00
29.6 - 30.4
(20)
31.50
9 31.2 - 31.8
(20)
5.0
8.0
12.6
The s i g n i f i c a n t d i f f e r e n c e s
an a s t e r i s k (*).
SILT
(P < 0.05)
39.50
39.2 - 39.8
(i0)
36.80
* 36.6 - 37.0
(i0)
38.60
* 38.1 - 39.1
(i0)
35.40
* 35.2 - 35.6
(i0)
are m a r k e d
with
The
thermal
preferences
of
ectothermic
organisms
c h a r a c t e r i z e t h e i r c a p a c i t y to r e s p o n d to the d i r e c t i n g
influence
of
temperature.
Reynolds
and
Casterlin
(1977) h a v e m e n t i o n e d t h a t fish, t o g e t h e r w i t h o t h e r
vertebrates,
exhibit
this
capacity
by
occupying
a
n a r r o w range w h e n p l a c e d in an ample t h e r m a l g r a d i e n t
in the laboratory. This is i n t e r p r e t e d as a r e f l e c t i o n
of
the
precision
of
thermoregulator
behavior.
Any
c h a n g e in the f a v o r i t e t e m p e r a t u r e of the fish serves
as an i n d i c a t o r of a l t e r a t i o n s c a u s e d by c h a n g e s in the
environment, such as thermal stress or the p r e s e n c e of
p e s t i c i d e s (Olgivie and A n d e r s o n 1965). This latter is
due to the p r e c i s i o n and s p e c i f i c i t y of t h e r m o s e l e c t i o n
in fish. The above m e n t i o n e d a u t h o r s d e m o n s t r a t e d that
salmon
(Salmo
salar)
selects
lower
or
higher
t e m p e r a t u r e s w h e n s u b j e c t e d to DDT d e p e n d i n g on w h e t h e r
the c o n c e n t r a t i o n of the c o n t a m i n a t i n g a g e n t is h i g h or
low. P e t e r s o n
(1973) p r o v e d t h a t S. s a l a r p r e f e r r e d
temperatures
5 to 7~
h i g h e r t h a n the c o n t r o l s w h e n
subjected
to the g r e a t e r
concentrations
of
several
pesticides, and o b s e r v e d s i m i l a r e f f e c t s in S a l v e l i n u s
fontinalis.
The
juveniles
of
the
herbivorous
carp
selected
temperatures
h i g h e r t h a n those at w h i c h the a n i m a l s
a c c l i m a t i z e d . The acute p r e f e r e n d u m that c h a r a c t e r i z e d
thermoselection
was 29~
The v a l u e of the s e l e c t e d
t e m p e r a t u r e i n c r e a s e d as the d e t e r g e n t c o n c e n t r a t i o n in
the e n v i r o n m e n t increased. A c o n c e n t r a t i o n of 12.6 m g / L
of a c t i v e agent b r o u g h t about s i g n i f i c a n t
(P < 0.05)
662
differences
with
respect
to
the
control.
This
is
T a b l e 2. G r o w t h rate (mg/day) of C. idella e x p o s e d to
different detergent concentration
for 21 days
(mean
values). N = n u m b e r of organisms.
Detergent
0.0
5.0
8.0
12.6
N
30
30
30
30
Initial w e i g h t
(g)
Final w e i g h t
(g)
1.35
1.68
1.69
1.34
/weight
(mg/day)
1.77
1.95
1.77
1.39
The s i g n i f i c a n t d i f f e r e n c e s
an a s t e r i s k (*).
(P < 0.05)
20.00
12.86 *
3.81 *
2.38 *
are m a r k e d w i t h
i n t e r p r e t e d as a s y m p t o m of stress since the p r e f e r r e d
t e m p e r a t u r e r e p r e s e n t s the thermal g r a d i e n t interval in
which
the
fish
carry
out
their
functions
more
e f f i c i e n t l y (Kellog and Gift 1983).
The d i f f e r e n c e was
only of 1.5~ but i n d i c a t e d thermal d i s o r i e n t a t i o n .
Fish g r o w t h rate was c a l c u l a t e d in order to e s t a b l i s h
the d e g r e e of stress caused by the detergent, as g r o w t h
r e s p o n d s s p e c i a l l y well to the s e l e c t e d temperature.
C o m p a r e d w i t h the control, a 36.9% d e c r e a s e in the
growth
rate
of
fish
subjected
to
the
lesser
c o n c e n t r a t i o n and a d e c r e a s e g r e a t e r than 88% in that
fish
subjected
to
the
greater
concentration
were
registered.
In a p r e v i o u s p a p e r (Espina et al. 1986)
the e n e r g y b a l a n c e of j u v e n i l e s of C. idella was found
to be a l t e r e d by alkyl aryl sulphonate.
The d i f f e r e n c e b e t w e e n the s e l e c t e d t e m p e r a t u r e and the
SILT is 9.5 ~ in the control groups of j u v e n i l e s of C.
idella
and it d e c r e a s e s to 5.8 C and to 3.9 C at the
lower and h i g h e r c o n c e n t r a t i o n of the c o n t a m i n a t i n g
agent respectively.
The effect of the d e t e r g e n t is
also e v i d e n t in the d e c r e a s e in the s u p e r i o r i n c i p i e n t
lethal temperature, and this d e c r e a s e d e p e n d s on the
c o n c e n t r a t i o n of the d e t e r g e n t in the environment.
O
O
--
The
exposure
of
the
organisms
to
sublethal
c o n c e n t r a t i o n s of d e t e r g e n t altered the r e s p o n s e s to
the d i r e c t i n g
influence
of t e m p e r a t u r e
as well
as
t e m p e r a t u r e acts as a m o d u l a t i n g factor.
Acknowledgments.
We
thank
the
Centro
Piscicola
T e z o n t e p e c de Aldama, H i d a l g o and Biol. G e r a r d o Cruz
for s u p p l y i n g C. idella juveniles.
The t r a n s l a t i o n
E n g l i s h w r i t i n g by Biol. Ma. A n t o n i e t a A r i z m e n d i
g r e a t l y appreciated.
663
de
O.
to
is
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664
Guille A