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Comparative Biochemistry and Physiol
Acute effects of sediments taken from an urban stream on physiological
and biochemical parameters of the neotropical fish Prochilodus lineatus
Juliana S. Almeida, Paulo C. Meletti, Claudia B.R. MartinezT
Departamento de Ciencias Fisiologicas, Universidade Estadual de Londrina, C.P. 6001 CEP: 86051-990, Londrina, Parana, Brazil
Received 28 October 2004; received in revised form 10 March 2005; accepted 14 March 2005
Abstract
Juveniles of Prochilodus lineatus were exposed to sediments collected from one of five sites along an urban stream into which various
types of contaminants are discharged. After 24 or 96 h fish were examined and the results compared with those from control groups (fish
exposed only to water, for the same period). Plasma ion levels varied significantly and fish exposed to site 5 sediment showed a transient
increase in both sodium and chloride concentrations. Plasma glucose was significantly higher in fish exposed to sediment from sites 2 and 5.
The higher liver glutathione-S-transferase activity registered in fish exposed to sediment from sites 1, 4 and 5 suggests the presence of
organic contaminants at these sites and the enhancement of liver catalase activity in fish exposed to sediment from sites 3 and 4 may be due to
contaminant-mediated oxyradical production. The overall results revealed that sites 4 and 5 are more severely contaminated, probably due to
organic contaminants from agricultural sources and municipal landfill.
D 2005 Elsevier Inc. All rights reserved.
Keywords: Fish; Sediment toxicity test; Biomarkers; Biological monitoring; Cambe stream; Plasma ions; Blood glucose; Liver catalase; Liver glutathione-S-
transferase; Hepatosomatic index
1. Introduction
It is important to monitor the aquatic environment,
especially sediments, which are sinks for many pollutants
(Lyytikainem et al., 2001). Sediments can accumulate
large quantities of chemicals, particularly poorly soluble
organic compounds, that may be rapidly taken up by
benthic fish, both through direct contact with the sediment
and interstitial water, and from ingested food (Vigano et
al., 2001). Thus, pollution monitoring or impact assess-
ment in aquatic ecosystems should not be limited to the
water phase, but should also include the sediment.
Historically, the assessment of sediment quality has often
been restricted to chemical analysis. However, quantifying
contaminant concentrations alone cannot provide enough
information to evaluate adequately the potential adverse
1532-0456/$ - see front matter D 2005 Elsevier Inc. All rights reserved.
doi:10.1016/j.cca.2005.03.004
T Corresponding author. Fax: +55 43 3371 4207.
E-mail address: cbueno@uel.br (C.B.R. Martinez).
effects, or time-dependent availability of these materials to
aquatic organisms (Ingersoll, 1995). Hence, the employ-
ment of sublethal sediment toxicity tests seems to be a
more effective means of predicting and detecting the
diverse impacts of contaminated sediments. In the last
decade fish have been increasingly used in toxicity tests
with sediment (Di Giulio et al., 1993; Vigano et al., 1998,
2001).
Tropical ecosystems are currently threatened by human
activities and environmental degradation; however, little
research has been done on the impact of contaminants on
tropical ecosystems and aquatic biota (Lacher and Gold-
stein, 1997) and not many tropical fish species have been
employed in toxicity tests. Thus, there is a real lack of data
concerning the effects of toxic agents on these fish species.
The neotropical freshwater fish Prochilodus lineatus
(Valenciennes, 1847) (=P. scrofa Steindachner, 1881)
represents a well suited species to sediment toxicity tests
due to its detritivorous habit, meaning that it can be in
contact with xenobiotics in sediment as well as in water
ogy, Part C 140 (2005) 356 – 363
J.S. Almeida et al. / Comparative Biochemistry and Physiology, Part C 140 (2005) 356–363 357
(Da Silva et al., 2004), and also because this fish has been
shown to be sensitive to toxicants (Mazon and Fernandes,
1999; Martinez and Souza, 2002; Martinez et al., 2004)
and is considered a potential vertebrate bioindicator for
environmental monitoring (Cerqueira and Fernandes,
2002).
The objective of this study was to evaluate the quality of
sediments sampled at five stations along a polluted stream,
in Southern Brazil, by measuring biochemical and physio-
logical variables of P. lineatus after exposure to the
sediments. The results of these analyses combined with
chemical data from the sampling sites were used to rank the
degree of contamination along the stream.
2. Materials and methods
2.1. Study sites and sample collection
The Cambe stream and its tributaries constitute the
main hydrological basin of Londrina, a city of 500,000
inhabitants in Parana state, Southern Brazil. This stream
crosses the entire city and receives diffuse and point
source discharges of mixed contaminants. Superficial
sediment samples were collected from the river-bed at
five sites along this stream (Fig. 1): site 1, into which
domestic sewage and mixed industrial effluent are dis-
charged; site 2, which receives waste from a car battery
factory; site 3, which is inside a Municipal Park and is
more preserved; site 4, located in an agricultural area, and
site 5, into which flows the leachate from the municipal
landfill and which is also situated in an agricultural area.
Sediment samples at each location were collected with a
Van Veen grab and stored in the dark, at 4 -C, for up to
15 days, as recommended by ASTM (1994). Water
samples were collected at the same sites and stored at
�20 -C for analysis of nutrients. Water temperature,
dissolved oxygen, pH and conductivity were measured at
the sampling sites.
Fig. 1. Location of the study area showing the sites of sediment
2.2. Metal and nutrient analysis in sediment and water
samples
Sediment samples were dried at 120 -C for 12 h. One
gram of dry sediment was placed in a 250 mL Erlenmeyer
flask and 25 mL of 0.1 M HCl solution was added. Samples
were shaken on a horizontal shaker for 2 h. The solution was
then filtered and analyzed for chromium, copper, zinc, lead
and manganese concentrations by atomic absorption spec-
trophotometry. Water samples were analyzed for ammonia
and nitrite (Golterman et al., 1978), nitrate and total
phosphorus (Mackereth et al., 1978) and total nitrogen
(Valderrama, 1981). Sediment and water from each site were
analyzed in triplicate.
2.3. Sediment-toxicity tests
Juveniles of P. lineatus (19.40T8.53 g, n =102), suppliedby the Universidade Estadual de Londrina Hatchery Station,
were held in a 600 L tank for at least 7 days with aerated
well-water prior to the experiment. Fish were fed with pellet
food each 48 h. Feeding was suspended 24 h before the
experiments began.
Short-term (24 and 96 h) static toxicity tests were
carried out to evaluate the toxicity of each sediment to P.
lineatus. Experiments were performed in 100 L glass
aquaria containing 6 fish each, with continuous aeration
and a photoperiod (L:D) of 14:10. Sediment samples were
homogenized, and 8 kg of sediment were placed in the
bottom of the aquarium, which was then slowly filled with
water (80 L). Aquaria were equilibrated to test conditions
for 24 h prior to the addition of test animals. Control
groups consisted of animals that were exposed only to
water. Water temperature, dissolved oxygen, pH and
conductivity were measured at the beginning and at the
end of each experiment. For all the treatments, water
temperature, dissolved oxygen, pH and conductivity were
in the range 19.5–23.3 -C, 5.3–7.7 mg O2/L, 7.6–8.1 and
107–180 AS/cm, respectively. After 24 and 96 h of
sampling along the Cambe stream (Parana State, Brazil).
Table 1
Mean levels of nutrients (Ag/L) measured in water and metals (mg/kg dry
weight) measured in sediment from sampled field sites
Site 1 Site 2 Site 3 Site 4 Site 5
Nutrients in water
NO2� 49.68 30.72 39.12 589.86 805.55
NO3� 659.16 521.74 685.55 1138.47 1407.63
NH3 93.61 238.19 130.92 1729.15 1073.85
Total N 1104.02 1038.79 1169.25 14874.07 9531.74
Total P 82.62 23.90 31.08 813.69 669.85
Metals in sediment
Cr 1.42 4.00 7.00 1.58 5.75
Cu 50.74 93.99 123.41 46.74 47.74
Zn 80.43 51.26 307.09 27.09 32.51
Pb 24.67 1173.33 101.67 4.42 1.25
J.S. Almeida et al. / Comparative Biochemistry and Physiology, Part C 140 (2005) 356–363358
exposure blood samples were taken from the caudal vein
by means of heparinized plastic syringes. Biometrics
measurements were made and subsequently fish were
killed by cervical section and their livers immediately
removed, weighed and stored frozen at �80 -C. Hepato-somatic index (HSI) was calculated for each fish as (liver
weight / fish body weight)�100.
2.4. Physiological and biochemical measurements
A small amount of blood was used for hemoglobin
determination by the cyanomethemoglobin method. Plasma
samples were obtained by blood centrifugation (3000�g
for 10 min) and samples were stored frozen (�20 -C) untilused for chemical analysis. Plasma Na+ and K+ were
measured in diluted samples (1:100) against known stand-
ards by flame photometry. Plasma chloride concentration
was determined by the thiocyanate method using a
commercial kit (Analisa, Brazil). Plasma glucose was
analyzed using a colorimetric commercial kit (GLUCOX
500-Doles Reagentes, Brazil) based on the glucose-oxidase
reaction. Total plasma lipids were determined by a
spectrophotometric enzymatic method using a commercial
Table 2
Plasma Na+, Cl� and K+ concentrations of P. lineatus exposed for 24 or 96 h to s
clean water (CTR)
Site Dt [Na+] (mM) [Cl�] (m
CTR EXP CTR
1 24 161.5T3.5 (6) 142.4T2.0 (4)T 95.1T
96 164.2T5.6 (5) 179.6T9.1 (4) 106.1T
2 24 171.8T5.4 (5) 177.7T6.6 (6) 108.0T
96 166.6T5.2 (5) 166.8T4.3 (3) 106.1T3 24 192.5T8.0 (6) 188.8T3.9 (6) 116.2T
96 166.3T3.3 (6) 171.9T2.4 (6) 120.2T
4 24 157.7T3.2 (6) 161.0T2.2 (6) 109.7T96 166.3T3.3 (6) 161.7T2.6 (6) 120.2T
5 24 157.7T3.2 (6) 175.4T6.2 (6)T 109.7T
96 185.4T4.6 (6) 175.8T5.5 (6) 122.4T
Values are meanTS.E. (n).
T Different from respective control ( P�0.05).
kit (Analisa, Brazil). Fish livers were homogenized in 10
volumes (w/v) of ice-cold 0.1 M K-phosphate buffer (pH
7.0) and centrifuged (14,000�g) for 20 min at 4 -C, toobtain the supernatant for glutathione-S-transferase (GST)
and catalase analyses. GST activity was determined as
described by Keen et al. (1976) using 1-chloro-2,4-
dinitrobenzene (CDNB) as substrate. The change in
absorbance was recorded at 340 nm and the enzyme
activity was calculated as nM CDNB conjugate formed/
min/mg protein using a molar extinction coefficient of 9.6
mM/cm. Catalase activity was estimated from the rate of
consumption of hydrogen peroxide levels (Beutler, 1975).
Change in absorbance was recorded at 240 nm and
enzyme activity was expressed as AM H2O2 consumed/
min/mg protein. Total plasma and liver proteins were
measured by the method of Lowry et al. (1951) with
bovine serum albumin as standard. All samples were
analyzed in duplicate.
2.5. Statistical analyses
For each variable, differences between experimental and
control treatments, at each exposure time (24 and 96 h) were
analyzed by Student’s t-test with a confidence interval of
5% (P <0.05).
3. Results
The concentrations of nutrient in water and metal in
sediment at each sampling site are presented in Table 1.
Nutrient concentrations increased below site 3 and were
strikingly higher in water from sites 4 and 5, compared with
sites 1, 2 and 3. Sediment samples from site 2 had a very
high lead concentration (1173.33 mg Pb/kg) while metal
concentrations in sediment samples from sites 1, 4 and 5
were low and very similar.
Plasma ion levels varied significantly, however few
consistent patterns became apparent (Table 2). Fish exposed
ediment samples from different sites along Cambe stream (EXP) or only to
M) [K+] (mM)
EXP CTR EXP
2.5 (5) 103.4T2.7 (6) 4.0T0,4 (6) 3.9T0.4 (6)
6.0 (6) 125.7T1.1 (6)T 5.0T0.6 (4) 5.9T0.4 (6)
3.5 (6) 120.9T1.6 (6)T 5.0T0.4 (6) 4.6T0.4 (6)
6.0 (6) 109.6T6.6 (6) 5.0T0.6 (4) 4.3T0.4 (6)
3.6 (5) 130.3T3.6 (6)T 4.5T0.2 (6) 3.9T0.2 (6)T2.9 (6) 121.2T1.2 (6) 4.4T0.1 (6) 3.4T0.2 (6)T3.8 (6) 125.8T4.1 (6)T 4.9T0.4 (6) 4.9T0.1 (6)
2.9 (6) 119.0T2.4 (6) 4.4T0.1 (6) 2.8T0.1 (6)T3.8 (6) 127.8T5.4 (6)T 4.9T0.3 (6) 4.7T0.1 (6)
3.9 (5) 126.7T3.3 (5) 5.0T0.3 (6) 4.8T0.2 (6)
Table 3
Plasma glucose, total lipid and total protein concentrations of P. lineatus exposed for 24 or 96 h to sediment samples from different sites along Cambe stream
(EXP) or only to clean water (CTR)
Site Dt Glucose (mg/dL) Lipids (mg/dL) Proteins (mg/mL)
CTR EXP CTR EXP CTR EXP
1 24 31.0T1.7 32.4T3.3 573.4T46.1 1087.2T124.0T 3.48T0.18 3.61T0.27
96 22.2T1.9 21.3T4.3 726.4T64.0 775.3T105.8 4.37T0.17 3.41T0.17T2 24 23.0T1.6 45.1T7.2T 727.3T106.1 786.2T38.2 3.63T0.18 3.79T0.20
96 22.2T1.9 19.9T1.4 726.4T63.9 568.4T46.7 4.37T0.17 3.68T0.15T3 24 28.5T3.7 31.9T6.6 681.8T71.2 702.0T60.0 3.35T0.17 3.16T0.17
96 27.0T3.3 21.3T4.7 1216.4T42.5 1093.23T85.3 3.91T0.11 4.04T0.074 24 31.1T2.7 33.4T1.8 1410.3T136.9 1582.1T81.4 3.35T0.21 3.81T0.12
96 27.0T2.7 26.5T3.4 1216.4T42.5 1143.2T73.3 3.91T0.11 3.20T0.09T5 24 31.1T3.3 30.7T3.6 1410.3T136.9 1092.6T165.1 3.35T0.21 2.43T0.25T
96 21.1T3.3 30.1T1.7T 738.3T125.1 905.8T46.6 2.46T0.19 2.90T0.10T
Values are meanTS.E. (n =6).
T Different from respective control ( P <0.05).
J.S. Almeida et al. / Comparative Biochemistry and Physiology, Part C 140 (2005) 356–363 359
to site 5 sediment showed a transient increase in both
sodium and chloride concentrations after 24 h, compared to
control (P <0.05) and a return to control values after 96 h.
Also, chloride levels increased, with no corresponding
variation in sodium, in fish exposed to sites 2, 3 and 4
sediment, for 24 h, and to site 1 sediment, for 96 h
(P <0.05). On the other hand, plasma potassium was lower
than control in animals exposed to site 3 sediment for both
experimental periods (P <0.05) and in animals exposed to
site 4 sediment, for 96 h (P <0.01).
Plasma glucose, a metabolic stress indicator, was
significantly higher (P <0.05) than control in fish exposed
to sites 2 and 5 sediment for 24 and 96 h, respectively (Table
3). Total lipids plasma concentration increased significantly
only in fish exposed to site 1 sediment, for 24 h (P <0.01).
P. lineatus exposed for 96 h to sites 1, 2 and 4 sediments
exhibited a significant decrease in plasma protein levels in
relation to their respective controls (P <0.05), whereas fish
exposed to site 5 sediment showed decreased plasma protein
24h 96h 24h 96h 24h 96h
0
0.2
0.4
0.6
0.8
HS
I
SITE 1 SITE 2 SITE 3
*
Fig. 2. Hepatosomatic index (HSI) of P. lineatus exposed for 24 or 96 h to sedimen
water (CTR). *Indicates values statistically different from respective controls ( P
levels after 24 h of exposure and increased protein
concentrations after 96 h in relation to their respective
controls (P <0.05).
P. lineatus exposed to sites 2 and 4 sediment for 24 h and
to site 5 sediment for both experimental periods showed
reduced values of HSI, but only the site 4 result was
statistically significant (P <0.01). On the other hand,
significant increases in HSI (P <0.01) were observed in
fish exposed to sites 3 and 4 sediment, after 96 h, compared
to control values (Fig. 2).
Hepatic GST and catalase activities in fish exposed to
sediments are presented in Fig. 3. Control values for GST
ranged from 81.60 to 130.76 nM/min/mg ptn (108.86T17.7;meanTS.D.). GST activity was significantly higher than
control values in P. lineatus exposed to site 1 sediment for
24 h (P <0.05) and in fish exposed to sites 4 and 5
sediments for 96 h (P <0.05). Control values for catalase
ranged from 15.69 to 32.99 AM/min/mg ptn (25.43T5.81;meanTS.D.). Catalase activity was significantly higher than
24h 96h 24h 96h
CTR
EXP
SITE 4 SITE 5
*
*
t samples from 5 different sites along Cambe stream (EXP) or only to clean
<0.05).
50
75
100
125
150
% o
f co
ntr
ol a
ctiv
ity
Site 1 Site 2 Site 3 Site 4 Site 5
GST
24h 96h
60
80
100
120
140
% o
f co
ntr
ol a
ctiv
ity
Site 1 Site 2 Site 3 Site 4 Site 5
Catalase
24h 96h
* *
*
*
* *
Fig. 3. Hepatic activity of glutathione transferase (GST) and catalase of P. lineatus exposed for 24 or 96 h to sediment samples from different sites along Cambe
stream. Results are expressed as % of control activity. *Indicates values significantly higher than respective controls ( P <0.05).
J.S. Almeida et al. / Comparative Biochemistry and Physiology, Part C 140 (2005) 356–363360
control values in animals exposed to site 3 sediment, for
both experimental periods (P <0.05) and exposed to site 4
sediment after 96 h (P <0.05).
4. Discussion
Sediments Quality Guidelines (SQG) are often helpful
when interpreting sediment chemical measurements and
biological effects. MacDonald et al. (2000) have elabo-
rated numerical SQG to assess sediment quality in
freshwater ecosystems by matching sediment chemistry
and toxicity data from field studies conducted across
United States. For each contaminant two SQG were
developed: a threshold effect concentration (TEC) below
which harmful effects are unlikely to be observed, and a
probable effect concentration (PEC), above which harmful
effects are likely to be observed. In the present study,
chromium concentration in sediments from all sampling
sites and zinc concentration in sediment from sites 1, 2, 4
and 5 were lower than the TEC (43.4 mg/kg; 121 mg/kg,
respectively). On the other hand, sediment copper
concentrations at all sites exceeded the TEC (31.6 mg/
kg), but not the PEC (149 mg/kg). Concerning lead,
sediment from site 2 exceeded PEC (128 mg/kg), while at
site 3 the concentration was lower, exceeding only the
TEC (35.8 mg/kg). These results suggest that the five
sites on the Cambe stream are exposed to different levels
of metal contamination and that sites 2 and 3, both of
which are downstream from a car battery factory, are
most contaminated.
On the other hand, at sites 4 and 5, complex mixtures
of organic contaminants from agricultural sources were
expected to be present. Leachate from the municipal
landfill also represents an important source of contam-
ination at site 5, contributing a high load of organic
matter and nitrogen. Thus, organic contaminants were
expected to be responsible for the main alterations
observed in fish exposed to sites 4 and 5 sediments,
although they were not measured in the present study.
However, in addition to the contaminants, the outcome
of the sediment toxicity test is affected by many factors,
J.S. Almeida et al. / Comparative Biochemistry and Physiology, Part C 140 (2005) 356–363 361
which makes the interpretation of the results compli-
cated. The observed adverse effects of sediments cannot
be directly and solely connected to any physicochemical
characteristic of sediment or to any single toxicant,
rather being the result of various factors (Lyytikainem
et al., 2001).
In general, plasma Na and Cl tend to be similarly affected
by waterborne toxicants (Pelgrom et al., 1995). However, in
this study plasma Cl concentrations were more strongly
affected in sediment-exposed fish, while plasma Na
concentrations were unchanged, except in fish exposed to
site 5 sediment for 24 h, where the rise in Na matched that in
Cl. These ion increases could be the result of plasma water
moving out of the circulation and into the tissues, as already
observed in acutely stressed freshwater fish (Wendelaar
Bonga, 1997). However, the fact that the plasma protein did
not rise, but actually fell in those fish where there was an
increase in Na and Cl, argues against a hemoconcentration
effect operating alone and may reflect the effect of
contaminants of site 5 sediment on branchial Na and Cl
influx activity (Rotland et al., 2001). Unlike the other
plasma ions, plasma K concentrations decreased in fish
exposed to site 3 and site 4 sediments. This may reflect an
increase in potassium influx to erythrocytes and/or other cell
types in response to some contaminant present in these
sediments.
The biological significance of elevated glucose concen-
trations in fish exposed to sites 2 and 5 sediments, as well as
elevated plasma lipids in fish exposed to site 1 sediment, is
probably related to an increased demand for metabolic
substrates after exposure to a stress stimulus. Mobilization
of energy stores is required to maintain homeostasis during
a chemical challenge. Serum glucose and lipid levels are
affected by storage tissue release and tissue uptake for
utilization. Thus, serum concentrations of energy-related
biomolecules may be useful biomarkers of toxicant stress
(Mayer et al., 1992; Lohner et al., 2001). Protein may also
be used as an energy source during severe stress. In this
study, plasma protein levels were generally lower in
sediment-exposed fish (Table 2), indicating a possible
nutritional imbalance.
The hepatic somatic index (HSI) can reflect both
metabolic energy demand and short-term nutritional status
and can be considered a general health indicator, sensitive
to environmental contaminants (Everaarts et al., 1993). In
this study, fish exposed to sediment from sites 2, 4 and 5
showed a reduction in HSI (Fig. 2), which can be
interpreted as reflecting a depletion of energy reserves
stored as liver glycogen. The significant increase in
plasma glucose shown by fish exposed to sediment from
site 2 for 24 h, and from site 5 for 96 h, reinforces this
hypothesis. In the large majority of cases, acute exposure
to pollutants has been found to increase blood glucose
and deplete liver glycogen (Vigano et al., 2001). Such a
depletion of glycogen reserves is generally attributed to
an increased energy demand associated with chemical-
induced stress, suggesting greater contamination at sites 2,
4 and 5.
On the other hand, HSI was higher in fish exposed to
sediment from sites 3 and 4 for 96 h. Enlargement of the
liver can be caused by hyperplasia or hypertrophy and may
increase its capacity to biotransform xenobiotics (Martin
and Black, 1998). Porter and Janz (2003) also observed
increased HSI in male sunfish (Lepomis megalotis)
collected from a stream in which treated municipal sewage
effluent was discharged. According to the authors, the
increased liver size could be attributed to hypertrophy of
liver for the removal of toxicants. In fact, in this study, fish
that presented higher HSI also showed increased hepatic
activity of catalase and GST.
Interactions of xenobiotics with numerous enzyme
systems, including detoxifying enzymes, have been shown
to result in a rise of reactive intermediates and reactive
oxygen species (Machala et al., 1997). Glutathione-S-
transferases (GST) are a group of enzymes that catalyze
the conjugation of reduced glutathione (GSH) with a
variety of electrophilic metabolites, and are involved in
the detoxification of both reactive intermediates and
oxygen radicals (Di Giulio et al., 1995). It has been
demonstrated that the activity of these enzymes may be
enhanced in the presence of polycyclic and polychlori-
nated hydrocarbons (Zhang et al., 1990) and even low
level organic contamination can lead to increased hepatic
GST activity (Machala et al., 1997). Thus, the stronger
GST activity noticed in fish exposed to sediments from
sites 1, 4 and 5 suggests the presence of organic
contaminants at these sites. High GST activity was also
reported by Beyer et al. (1996) and Stien et al. (1998) in
fish caged in polluted places. These authors suggested
that the enzyme was induced by organic contaminants,
such as petroleum hydrocarbons and polychlorinated
biphenyls (PCBs).
Antioxidant enzymes, including catalase, are inducible
by conditions that increase the flux of reactive oxygen
species, such as O2�, H2O2 and OH
� (Di Giulio et al., 1993).
Much evidence indicates that xenobiotics can generate
reactive oxygen species, including hydrogen peroxide
(Wilhelm Filho et al., 2001). In this study, the enhancement
of catalase activity in fish exposed to sediment from sites 3
and 4 may be due to contaminant-mediated oxyradical
production. Increased liver catalase activity has been
observed in catfish exposed to sediment contaminated with
hydrocarbons (Di Giulio et al., 1993) and other fish species
from polluted sites (Livingstone et al., 1995; Wilhelm Filho
et al., 2001).
The present study showed that the use of a set of
non-specific biomarkers which respond to several types
of pollutants and are related to some key biochemical
and physiological functions such as detoxification
(GST), antioxidant defense (catalase), osmoregulation
(plasma ions) and metabolism (plasma glucose, lipids
and protein and HSI) represents a sensitive and effective
J.S. Almeida et al. / Comparative Biochemistry and Physiology, Part C 140 (2005) 356–363362
tool for sediment bioassays. The observed effects of
sediments on the selected biomarkers of P. lineatus made
it possible to rank the risk associated with different sites
on the Cambe stream and reveal that sites 4 and 5 are
the most severely contaminated. The sediment toxicity
tests using young P. lineatus have shown an interesting
potential for identifying the presence of toxicological risk
for the fish community, discriminating between poten-
tially threatened and unthreatened sites of the Cambe
stream.
Acknowledgments
The authors thank the Universidade Estadual de Lon-
drina hatchery station (EPUEL) for the supply of fish. They
are grateful to Indianara F. Barcarolli and Lıgia M. B.
Santana for significant field and laboratory assistance.
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