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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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