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Environmental Pollution 154 (2008) 68e76

www.elsevier.com/locate/envpol

Methylmercury in a predatory fish (Cichla spp.)

inhabiting the Brazilian Amazon
Helena do A. Kehrig a,*, Bruce M. Howard b, Olaf Malm a
a

Laboratorio de Radioisotopos Eduardo Penna Franca, Instituto de Biofsica Carlos Chagas Filho,
Universidade Federal do Rio de Janeiro (UFRJ), 21941-902 Rio de Janeiro, Brazil
b
T.H. Huxley School, Imperial College, London SW7 2BP, UK
Received 15 December 2007; accepted 21 December 2007

Cichla spp. may be considered good bioindicators of methylmercury contamination in

the Amazonian ecosystem because of their integration of this pollutant over time.
Abstract
This research tested whether limnological conditions, biological characteristics of fish and anthropogenic impacts influenced the assimilation
of methylmercury into the muscle of a sedentary piscivorous fish, Cichla spp., from three rivers (Negro, Madeira, Tapajos) and two hydroelectric
reservoirs (Balbina, Tucuru) within the Brazilian Amazon. Methylmercury in this fish ranged from 0.04 to 1.43 mg g1 w.w. across sites. No
significant differences were observed in the methylmercury concentrations between males and females, or for different morphotypes of this species. Positive correlations were found between methylmercury and fish body weight. No differences were found between the weight normalized
methylmercury (MeHg) concentrations or its percent of total mercury in fish from the three rivers; weight normalized MeHg was highest in one
of the two reservoirs. In Rio Tapajos, where gold mining and deforestation cause high water turbidity, fish showed the highest MeHg and
concentrations were different across the four sites examined. In all sampling areas, the %MeHg was found to be higher than 70.
2007 Elsevier Ltd. All rights reserved.
Keywords: Methylmercury; Predatory fish; Amazon tributaries; Hydroelectric reservoirs

1. Introduction
The concentrations of mercury in fish of Amazonian rivers
have been a matter of concern for health of humans and wildlife for over 15 years. Deforestation for agricultural projects,
damming for hydroelectric power plants, and alluvial gold
extraction in the Amazonia are increasing mercury discharges
in aquatic systems, exposing riverine populations to organic
mercury (methylmercury) through the consumption of fish
(Dolbec et al., 2000; Dorea and Barbosa, 2007). Mercury
* Corresponding author. Laboratorio de Radioisotopos Eduardo Penna
Franca, Instituto de Biofsica Carlos Chagas Filho, Universidade Federal do
Rio de Janeiro (UFRJ), Av. Carlos Chagas Filho, 373, 21941-902 Rio de Janeiro, Brazil. Tel.: 55 21 2561 5339; fax: 55 21 2280 8193.
E-mail address: kehrig@biof.ufrj.br (H.do A. Kehrig).
0269-7491/$ - see front matter 2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.envpol.2007.12.038

concentrations in forest soils were 1.5e3.0 times higher than

in pasture soils, suggesting strong re-mobilization after
deforestation (Lacerda et al., 2004). Mercury concentrations
in many Amazonian soils are naturally high and mercury
released from soils into river system is thought to be higher
than the combined input from anthropogenic sources (Roulet
et al., 1998; Fadini and Jardim, 2001).
The interpretation of mercury data in fish depends on
knowledge of the sources and behavior of this element, including its methylation. There are two main known sources of
mercury that may lead to mercury accumulation in fish from
the Amazon basin. The first and most widely known is artisanal gold mining that involves use of metallic mercury
(Hg ) to form an amalgam with the fine gold particles in soils
and sediments of some parts of the river. It has lead to the
discharge of mercury directly into rivers and emission to the

H.do A. Kehrig et al. / Environmental Pollution 154 (2008) 68e76

we examined whether Cichla spp. from sites with different

limnological habitats and anthropogenic activities (deforestation for agricultural projects, damming for a hydroelectric
power plant, and alluvial gold extraction) varied in their methylmercury concentrations and whether biological characteristics of these fish (sex, body weight, standard length and
different morphotypes of Cichla species) affected their
mercury concentrations.
2. Materials and methods
2.1. Sampling
In 1994 and 2000, specimens of adult Cichla spp. (tucunare) were collected by Laboratorio de Radioisotopos of the Federal University of Rio
de Janeiro (LREPF) at the three major tributaries of Rio Amazonas (Tapajos,
Madeira and Negro) and from two reservoirs (Balbina, Tucuru) within Brazilian Amazon (Fig. 1). Tables 1e3 show the number of fish collected at
each site (N) and the corresponding sampling year. Seasonal variations in
fish mercury concentrations have not been observed in the main river systems (Malm, personal communication). The exception being the case of Tucuru collected from another man made reservoir (Malm et al., 2004) and
from Balbina (Malm, personal communication).
The fish were caught with nets or obtained from local fishermen who use
a variety of fishing techniques. Following an examination of the visual characteristics of the samples collected from each area and comparison with the literature, it was concluded that at least four species were present in the samples
obtained: Cichla monoculus, Cichla temensis, Cichla ocellaris and Cichla sp.
An important limitation in the identification of species in any one location is

Cichla spp

Venezuela
Colombia
6

1
a

Manaus
Santarm
a Curi (Brasilia Legal)
b Itaituba
c Jacareacanga
1 Rio Amazonas
2 Rio Tapajs
3 Rio Negro
4 Rio Madeira
5 Tucuru Reservoir
6 Balbina Reservoir

Atlantic Ocean

Brazil

Argentina

Chile

Para

y
gua

Bolivia

Pacific Ocean

atmosphere as a result of heating the amalgam to recover the

gold. However, although gold mining activities may result in
local contamination around the mining centers, erosion of soils
as a result of human colonization may be another cause of
increased loading of particulate mercury into adjacent river
systems (Roulet et al., 1999). The mineral horizons of feralsols in the Amazonian region constitute a natural mercury
reservoir that is greater than others worldwide (Roulet et al.,
1999). In addition to agriculture, gold mining may be an efficient way by which soils are released to the aquatic system. In
addition to occupational exposure with the deforestation, since
the late 1980s the artisanal gold mining has caused serious
contamination of Amazonian waters. However, high total
mercury concentrations in fish and human hair have also
been observed in areas far removed from mines (Kehrig
et al., 1998; Dorea, 2003; Belger and Forsberg, 2006; Bastos
et al., 2007; Dorea and Barbosa, 2007).
There is a need for a basin-wide and systematic approach to
sampling and analysis of mercury in fishes from different
tributaries and reservoirs in the Brazilian Amazon. This means
selecting species (or possibly genera) that are representative of
the location from which they are captured, and paying attention to factors that are related to the variation in the percentage
of total mercury that is present as methylmercury. The present
study makes an initial contribution to this task by presenting
methylmercury data from three major rivers and two reservoirs
within Brazilian Amazon. The present manuscript adds new
insight to existing studies and also complements and compares
their findings to data generated by this study.
The sedentary piscivorous fish species, Cichla spp.
(tucunare), is useful for this purpose because it is commonly
consumed, easily identified (at least to the level of genus)
and may be considered good bioindicators of mercury in the
Amazonian ecosystem, especially because they integrate mercury over time. The genus Cichla is comprised of 15 nominal
species recognized by external characters, of which color pattern and meristics are most significant (Kullander and Ferreira,
2006). Their preferred habitat is lentic (slow moving) water.
Cichla are exclusively piscivorous and opportunistic in their
feeding habits. The maturation of Cichla for both sexes occurs
at approximately 20e35 cm or one to two years of age. This
discrepancy is likely to account for differences in the definitions of ripe gonads (Howard, 2001). Cichla is widely distributed in the Rio Amazonas basin (Kullander and Ferreira,
2006). This fish is an important food fish in the diet of some
Amazonian human populations, being consumed several times
a week in several riverine communities. Cichla presents a risk
of potentially harmful methylmercury exposure to humans. Its
commerce is an important economic activity throughout the
region.
The present study was aimed at answering the following
question: do different limnological and biological characteristics and human activities influence the assimilation of methylmercury by the muscle of a sedentary piscivorous fish species
Cichla spp. from three important rivers basins (Negro,
Madeira and Tapajos) and two hydroelectric reservoirs
(Balbina and Tucuru) within Brazilian Amazon? In this study

69

1250 km

Fig. 1. Sampling areas in the Brazilian Amazonian region.

H.do A. Kehrig et al. / Environmental Pollution 154 (2008) 68e76

Relation MeHg and L

log MeHg  1.1 0.02 * L

r2 0.50; p < 0.05
MeHg  0.06 0.01 * L
r2 0.10; p > 0.05
MeHg  0.2 0.02 * L
r2 0.24; p < 0.001

log MeHg 1.2 0.02 * L

r2 0.20; p 0.05
MeHg  0.2 0.03 * L
r2 0.33; p < 0.01

Relation MeHg and W

log MeHg  2.3 0.7 log W

r2 0.45; p < 0.05
log MeHg  1.1 0.3 log W
r2 0.14; p > 0.05
log MeHg  2.4 0.7 log W
r2 0.37; p < 0.0001

log MeHg  2.1 0.6 log W

r2 0.19; p 0.05
log MeHg  1.1 0.3 log W
r2 0.23; p < 0.01

log W  0.6 2.1 * log L

r2 0.63; p < 0.05
log W  1.8 2.9 * log L
r2 0.74; p < 0.05
log W  1.3 2.7 * log L
r2 0.82; p < 0.0001

log W  0.1 1.9 * log L

r2 0.78; p < 0.0001
log W  2.3 3.4 * log L
r2 0.96; p < 0.0001

Total mercury (Hg) concentrations in the wet muscle of fish were determined by cold vapor atomic absorption spectrometry with a Flow Injection
Mercury System (FIMS) e FIAS 400 (Perkin Elmer, USA) equipped with
auto sampler AS90 (Perkin Elmer, USA) and using sodium borohydride as a reducing agent (Kehrig et al., 2006).
The methylmercury (MeHg) analysis in the muscle was made by digesting
samples with an alcoholic potassium hydroxide solution followed by dithizone-toluene extraction. After series of clean-up steps, MeHg dithizonate
was identified and quantified in the toluene layer on a Shimadzu gas chromatograph GC-14 with an electron-capture detector-ECD (Kehrig and Malm,
1999). The detection limit for methylmercury in fish was 0.0005 mg g1 wet
weight (w.w.) for an average 0.5 g sample.
Analytical quality for total mercury and methylmercury was assured by
means of inter-laboratory comparison exercises between the LREPF and the
National Institute for Minamata Disease (NIMD) laboratories using fish samples from the Balbina Reservoir. Methylmercury results for the analysis performed at the two laboratories for 32 fish samples exhibited highly
significant coefficients of correlation (r2 0.99). The gradient of the regression line representing the relationship between analyses in the two laboratories
was close to 1 (slope, b 1.05).
Certified reference materials from International Atomic Energy Agency e
Monaco (IAEA-350) and from National Research Council-Canada (Dorm-2)
were analyzed in all sample batches. Our total mercury (Hg) and methylmercury (MeHg) analyses of certified reference samples analysis demonstrated the
high precision and accuracy of the analytical methods. Total mercury and
methylmercury quantified in the reference materials were within 95 and
98% of the mean certified values. The overall reproducibility for the analysis
period was determined from the results obtained using certified samples. The
coefficient of variation (SD/mean) for the duplicate samples was less than
10%.

18e37
108e985
19
Tucuru reservoirc (2000)

C. ocellaris (19)

17
Balbina reservoirc (1995)

2.2. Chemical analysis

N corresponds to the number of fish specimens collected. F, female; M, male.

a
Mining impacted area.
b
Not identified.
c
Not directly mercury impact.

17e51.5
97e740

F (10)
M (9)

25.5e56
250e4800
42
Rio Tapajosa (1998)

C. monoculus (18)
C. temensis (5)
Cichla sp. (19)
C. ocellaris (17)

32e38
350e570
5
Rio Negroc (1994)

F (9)
M (10)

23.5e46
216e937

C. monoculus (4)
C. temensis (4)
C. monoculus (5)
a

Rio Madeira (1994)

minemax
minemax

Standard length
(cm) (L)
Morphotypes of
Cichla species
N

Sex

Body weight
(g) (W)

Relation W and L

the potential for hybridization; so-called species may best be regarded as

end points.
Following determination of the body weight, standard length and identification of the species of Cichla (when possible), a skinless cube of the white
dorsal muscle tissue was extracted. Cubes of tissue were stored in airtight plastic bags at below 10  C until analysis. A total of 91 samples of fish specimens of Cichla spp. were analyzed for total mercury and methylmercury at
LREFP.

Sampling area
(sampling year)

Table 1
Biological characteristics of a sedentary piscivorous fish species, Cichla spp., and regression relationships between body weight and standard length, and between methylmercury concentrations (mg g1 w.w.) in
muscle and weight or standard length of the fish

70

2.3. Normalization of the methylmercury data

Methylmercury concentrations in the Cichla spp. muscle were subjected to
weight normalization in order to compare their levels within and among rivers
and reservoirs, due to the different mean body weights and standard lengths of
the fish collected at the various locations (see Tables 1 and 3).
The MeHg concentrations are best standardized to the length of the fish,
since length does not decrease with time while weight sometimes does (Huchabee et al., 1979). However, a non-significant relationship ( p 0.05) was
found between the standard length of the 91 specimens of Cichla spp. and
the concentrations of methylmercury. In this case, we normalized the concentrations of MeHg using the body weight of Cichla spp., using an average fish
weight, and computed the MeHg concentration from the logelog regression
equation (log MeHg  1.86 0.5 log W). This logelog regression equation
presented a significant relationship (r2 0.59, p < 0.0001) between concentrations of MeHg and body weight (W) of the 91 specimens of Cichla spp.

2.4. Statistical analyses

Statistical analyses were performed using STATISTICA 6.0 for Windows
(StatSoft, Inc 1984e2001, USA). After verification of the non-normal distribution of each data set, an analysis of variance was done by KruskaleWallis ANOVA followed by a Post hoc test (ManneWhitney U-test) to compare the
concentrations of MeHg normalized to a standard weight or the percent
MeHg of total Hg in muscle tissue of Cichla spp. from the three rivers and

H.do A. Kehrig et al. / Environmental Pollution 154 (2008) 68e76

71

Table 2
The range and mean of methylmercury concentrations (mg g1 w.w.), the mean methylmercury concentrations normalized to a standard body weight, and percent
methylmercury to total mercury in Cichla spp. from three rivers and two reservoirs of the Amazon Basin
N

Sampling area
(sampling year)

Rio Madeira (1994)

Rio Negrob (1994)
Rio Tapajosa (1998)
Balbina reservoirb (1995)
Tucuru reservoirb (2000)
a
b

8
5
42
17
19

Methylmercury concentration
(mg g1 w.w.)

Methylmercury concentration
normalized at standard
body weight (mg g1 w.w.)

Percent methylmercury
of total mercury

Average  SD (minemax)

Average  SD

minemax

0.47  0.29
0.38  0.06
0.55  0.38
0.24  0.16
0.49  0.27

0.63  0.22
0.61  0.12
0.48  0.34
0.48  0.37
1.12  0.78

82e102
74e99
61e104
72e103
71e106

(0.27e1.10)
(0.32e0.58)
(0.087e1.43)
(0.070e0.72)
(0.20e1.09)

Mining impacted area.

Not directly mercury impact.

two reservoirs or between the different morphotypes of Cichla species collected at each sampling area. A linear regression was performed to determine
the relationship between the biological parameters (weight or standard length)
and the concentrations of MeHg in muscle tissue. Values are presented as
mean  standard deviation (SD) based on wet weight (w.w.).

3. Results and discussion

Piscivorous species, which are at the top of aquatic food
webs, are a good indicator of mercury contamination in fish.
Amongst the data generated as part of the present study,
only 13 samples (14%) of the samples of Cichla spp. muscle
presented a total mercury concentration higher than the maximum permissible limit of 1.0 mg g1 w.w. established for human consumption of predatory fish in Brazil (Brasil, 1998).
Meanwhile, 31 fish samples analyzed (34%) had MeHg concentrations above the maximum limited of 0.5 mg Hg g1
w.w. established for food by the WHO. This does not mean,
however, that there is lower risk to human health, as the early
signs of the neurotoxic effects that could be related to MeHg
exposure from fish consumption have been shown in at least
one riverine community in the Rio Tapajos basin (Lebel
et al., 1998; Grandjean et al., 1999; Dolbec et al., 2000).
Some biological characteristics of Cichla spp. (sex,
morphotype of Cichla species, range of fish body weight and
standard length) from three rivers and two reservoirs of the
Amazon Basin are presented in the Table 1. Mean concentrations, on a wet weight basis, of MeHg and the percent MeHg

to total Hg in the muscle samples of Cichla spp. are presented

in Tables 2 and 3.
Total Hg and MeHg concentrations in the 91 muscle
samples of Cichla spp. ranged from 0.05 to 1.57 mg Hg g1
w.w. and from 0.04 to 1.43 mg MeHg g1 w.w. In this study,
total Hg and MeHg concentrations found in the muscle samples of Cichla spp. were of the same order of magnitude as
those reported in earlier studies with these piscivorous species
from the three rivers, Madeira, Tapajos and Negro, and
Balbina reservoir (Kehrig and Malm, 1999; Santos et al.,
2000; Uryu et al., 2001; Barbosa et al., 2003; Belger and
Forsberg, 2006; Bastos et al., 2007; Dorea and Barbosa, 2007).
In most cases, the ratio of MeHg to total Hg was found to
be greater than 70%, indicating that organic mercury was the
predominant form in the fish muscle (Tables 2 and 3). This
pattern was also observed in previous studies with freshwater
predator fishes from Brazilian Amazon and worldwide (Akagi
et al., 1995; Kehrig and Malm, 1999; Mason et al., 2000;
Bowles et al., 2001; Horvat et al., 2003).
Methylmercury concentrations found in muscle samples of
Cichla spp. were variable within any one location (Table 2).
For instance, at Balbina and Tucuru reservoirs this variation
was over ten-fold. This variation is probably due to the large
range in fish weights (Table 1) and also to the abundance
and assortment of prey items for this species, which vary
with the hydrological cycles in the Amazonian ecosystem. In
a previous study, the fish species at the top of the food web
of Rio Madeira and Rio Negro, Cichla spp. and Hoplias malabaricus, also exhibited a wide range of mercury concentrations

Table 3
The range of fish body weight and standard length, the range and mean methylmercury concentrations (mg g1 w.w.), the mean of methylmercury concentration at
a standard body weight, and percent methylmercury to total mercury in Cichla spp. from four sampling points within the Rio Tapajos, Brazilian Amazon
Sampling Areas
in Rio Tapajos

Santarem
Lago Curia (Braslia Legal)
Itaitubab
Jacareacangab
a
b

8
10
8
16

Not directly mercury impact.

Mining impacted area.

Body weight
(g)

Standard length
(cm)

Methylmercury concentration
(mg g1 w.w.)

Methylmercury concentration
normalized at standard
weight (mg g1 w.w.)

Percent methylmercury
of total mercury

minemax

minemax

Average  SD (minemax)

Average  SD

(minemax)

250e4800
390e3100
400e2050
420e1750

27e61
28e55
33e56
26e44

0.16  0.10
0.50  0.37
0.89  0.39
0.62  0.34

0.23  0.15
0.40  0.33
0.62  0.29
0.57  0.36

76e92
61e104
81e96
72e104

(0.038e0.36)
(0.19e1.43)
(0.44e1.25)
(0.18e1.30)

72

H.do A. Kehrig et al. / Environmental Pollution 154 (2008) 68e76

(Dorea and Barbosa, 2007). According to Dorea and Barbosa

(2007), a wide variation in fish mercury bioaccumulation is
expected in the complex and biologically dense environment
of the Amazon rain forest.
Dietary exposure is likely to be an important factor explaining the differences in accumulation of Hg as MeHg in Cichla
spp. in this study. This genus is opportunistic in terms of prey
selection. Consequently, studies of diet composition in one location are not necessarily directly applicable to another. Little
is known regarding Hg accumulation in fish as a function of
hydrological cycles in the Amazonian ecosystem. Fish feeding
behavior adapts to changes in habitat due to seasonal inundation of the forest (Bastos et al., 2007). Dorea et al. (2006)
observed that changes in the aquatic environment brought by
seasonal inundation and hydrological cycles could be affecting
the bioaccumulation of mercury in different fish species from
the Negro River, Amazon. However, the bioaccumulation of
mercury during high and low waters were similar for Cichla
spp. from the Rio Madeira (Bastos et al., 2007).
3.1. Biological characteristics (sex, body weight,
standard length, and different morphotypes)
Significant relationships were found between logtransformed body weight (W) of Cichla spp. and their logtransformed standard length (L) ( p < 0.05; see Table 1).
To evaluate the body weight of Cichla from the three rivers
(Rio Madeira, Rio Negro and Rio Tapajos) and two reservoirs
(Balbina and Tucuru), we used the relationships between
body weight and standard length of Cichla spp. (Table 1) to
calculate a mass at the overall mean standard length of
33 cm. The body weight of approximately 640 g and 772 g
calculated for Cichla of Balbina and Tucuru reservoirs were
higher than the estimated values of 388 g, 480 g and 402 g
for fish from the rivers, Madeira, Tapajos and Negro, respectively. The highest body weight found in Cichla of Balbina
and Tucuru reservoirs could be because reservoirs are more
productive ecosystems, with high phytoplankton densities
and also higher nutrients concentrations (C, N, P) than in
rivers. In reservoirs, the slow moving waters favor a permanence of microorganisms in the water column that associated
with a high accumulation of organic matter, result in high
primary production waters and more prey items for fish.
Significant relationships were found between methylmercury concentrations in Cichla spp. and their body weight
(log-transformed) or standard length ( p < 0.05) (see Table
1). Probably, these relationships are related to the fact that
the diet of Cichla progresses from the consumption of algae
and alevins, to shrimp, and then to a diet of fishes (Howard,
2001); this species is planktivorous in the beginning, then
carnivorous and, in the mature stage, piscivorous. Cichla
capture larger, likely more contaminated prey items as they
grow, i.e. with increasing standard length.
Differences in growth rate between fish populations in this
study are a potential explanation for variations in MeHg concentrations. According to Jepsen et al. (1999), members of the
same Cichla species collected from different water bodies with

different hydrographic characteristics would exhibit variable

growth rates. Fish growth rates may affect mercury accumulation by changing the balance between the rate of deposition of
new tissue and the gut adsorption efficiencies for both nutrients and MeHg (Howard, 2001).
Methylmercury concentrations in the Cichla spp. muscle
were subjected to weight normalization in order to compare
their levels within and among rivers and reservoirs. Concentrations of total Hg and MeHg at a standard size were also
reported by Sorensen et al. (1990) and Kehrig et al. (2001).
The range and mean of weight-normalized concentrations of
MeHg are summarized in Tables 2 and 3. The analysis of
variance (KruskaleWallis ANOVA) analysis showed no
significant differences (N 91; H 6.74; p > 0.05) for the
normalized concentrations of MeHg among the different
morphotypes of Cichla species (Cichla monocullus, Cichla
ocellaris, Cichla temensis and Cichla sp (Table 1)). However,
the analysis of variance showed a significant difference
(N 42; H 9.04; p < 0.01) for the normalized concentrations of MeHg among the different morphotypes of Cichla
species (Cichla monocullus, Cichla temensis and Cichla sp.)
collected at Rio Tapajos.
A Post hoc test (ManneWhitney U-test) analysis showed
no significant difference (N 23; Z  0.59; p > 0.05) for
the normalized concentrations of MeHg between the different
morphotypes of Cichla species (Cichla monocullus and Cichla
temensis) collected at Rio Tapajos. However, the U test analysis revealed significant differences (Z 2.28 and 2.24;
p < 0.05) for the normalized concentrations of MeHg between
the different morphotypes of Cichla species, Cichla temensis
versus Cichla sp., and Cichla monocullus versus Cichla sp.,
respectively. Probably, these differences may be that the
individuals belong to two distinct genetic populations, which
have different nutritional, developmental and reproductive
characteristics.
In Rio Tapajos it is shown that, although differences
between morphotypes of Cichla were apparent, there was no
detectable variation between males and females. The Post
hoc test revealed no sex-related differences (N 19;
Z 0.25; p > 0.10) for the normalized concentrations of
MeHg.
The Post hoc test showed no significant difference (N 8;
Z  1.15; p > 0.10) for the normalized concentrations of
MeHg between the different morphotypes of Cichla species
(Cichla monocullus and Cichla temensis) collected at Rio
Madeira. The apparent similarity in mean normalized concentrations of MeHg between Cichla monocullus and Cichla
temensis are probably related to the fact that they have similar
feeding habits in all stages of life. According to Howard
(2001), both genera contain comparable concentrations of
Hg in muscle tissue; MeHg accumulation in Cichla, a sedentary fish, is more sensitive to the characteristics of the location
from which it was caught than migratory fish species.
In the Tucurui reservoir, where only one morphotype of
Cichla was sampled (Table 1 Cichla ocellaris), there was no
detectable differences between males and females. The Post
hoc test has shown no sex-related differences (N 19;

H.do A. Kehrig et al. / Environmental Pollution 154 (2008) 68e76

Z 0.25; p > 0.10) for the normalized concentrations of

MeHg in the Cichla spp. sampled in Tucuru Reservoir.
3.2. Impacts of different limnological habitats and
human activities
The three major tributaries of Rio Amazonas, Rio Negro,
Rio Madeira and Rio Tapajos, which present contrasting physical, chemical and biological characteristics, and the two
hydroelectric reservoirs, Balbina and Tucuru, are subject to
different mercury inputs (atmospheric inputs, gold mining extraction, agriculture, forestry and forest burning) and particular
biogeochemical characteristics. The analysis of variance
revealed significant differences within and among rivers (Rio
Tapajos, Rio Madeira and Rio Negro) and reservoirs (Balbina
and Tucuru) of the Brazilian Amazon for the normalized
concentrations of total Hg (N 91; H 16.55; p < 0.01)
and MeHg in the Cichla spp. (H 20.37; p < 0.0001). We
also found a significant difference (N 91; H 12.41;
p < 0.01) in the percent MeHg to total mercury within and
among rivers and reservoirs of Brazilian Amazon.
3.3. Rio Negro, Rio Madeira and Rio Tapajos
The analysis of variance revealed no significant differences
for the normalized concentrations of total Hg and MeHg
(N 55; H 4.03; p > 0.10 and H 6.01; p > 0.05, respectively) between Cichla spp. collected at the Rio Madeira, Rio
Negro and Rio Tapajos. In addition, the analysis of variance
has shown no significant difference (N 55; H 3.14;
p > 0.10) for the percent MeHg to total Hg between the three
major tributaries of Rio Amazonas (Tapajos, Madeira and
Negro).
In this study, normalized concentrations of MeHg in Cichla
spp. from Rio Negro, which is less affected by human activities and has no history of gold mining, were similar to those
found in the specimens from rivers greatly impacted by deforestation and agriculture and also, with a history of gold-mining
activity, as the Rio Tapajos and Rio Madeira (Table 2). However, our results contrast with those of Dorea and Barbosa
(2007); they found that Cichla spp. with similar weights
from the Rio Madeira had higher total mercury concentrations
than individuals from the Rio Negro.
In this study, MeHg concentrations and percent MeHg to
total Hg in the muscle samples of Cichla collected at the
Rio Negro, a black water river, may be reflecting the natural
background of mercury present in its ecosystem basin, amplified by its unique biogeochemical characteristics. The black
water characteristics of this river, in particular the dissolved
organic carbon concentrations and low pH (Silva-Forsberg
et al., 1999) are conducive to the methylation of mercury
and its effective bioaccumulation. Fadini and Jardim (2001)
studied the physicochemical characteristics of the Rio Negro
and concluded that 99.74% of its mercury was from natural
sources. Mercury concentrations in rivers and lakes waters,
as well as in soils and bulk precipitation were high,

73

considering the scarcity of anthropogenic point sources in

the Rio Negro basin (Fadini and Jardim, 2001).
Previous studies in Brazilian Amazonian have shown that
piscivorous fish sampled in different tributaries of the Rio
Amazonas far removed from gold-mining had the highest
concentrations of mercury (Barbosa et al., 2003; Belger and
Forsberg, 2006; Dorea et al., 2006). In contrast, Boudou
et al. (2005) compared the concentrations of mercury between
rivers of Amazonian French Guyana and reported no significant difference in fish mercury concentrations between rivers
with and without alluvial gold extraction. Despite, the Rio Madeiras higher sediment load as well environmental impacts
(deforestation, agriculture, hydroelectric reservoir, and alluvial
gold mining) on natural mercury release (Dorea and Barbosa,
2007), concentrations of methylmercury in Cichla collected in
this study were not different from the Rio Tapajos, a clear
water river in Amazonian ecosystem, also impacted by gold
extraction, deforestation and agriculture. According to Bastos
et al. (2006), a recent survey of fish mercury showed that,
notwithstanding the reduction of mercury emissions to the
Rio Madeira basin from gold mining activities, concentrations
in fish were similar to those measured during the gold rush, in
the late 1980s. Oliveira et al. (2001) showed that the upper
horizon of the Amazon soil receives an important mercury
contribution from anthropogenic activity. Anthropogenic
activities related to the fire deforestation, agriculture, and
gold mining can greatly increase mercury release from soil
and sediment (Dorea et al., 2004). Remobilization of mercury
from bottom sediments plus re-emission from soils due to land
use changes probably responsible for keeping high mercury
concentrations in these fish.
3.3.1. Rio Tapajos
The Rio Tapajos, one of the principal tributaries of the Rio
Amazonas, is one of great importance due to the fact that it
receives a large mercury inputs from anthropogenic activity,
and the basin also contributes half of the gold produced by
the gold mining areas in Brazil (Santos et al., 2000). In the
twentieth century, this river suffered the biggest impacts of
the gold mining activities. Despite, the Rio Tapajos is a clear
water river, in areas close to the goldmine activities; the river
waters have high amounts of suspended particulate matter rich
with mercury (Howard, 2001). This is particularly the case
along the main channel of the Rio Tapajos from Jacareacanga
to Itaituba cities, where the intense gold mining centre
increased the turbidity of the water (Howard, 2001).
However, Roulet et al. (2001) showed that the dominant
source of mercury in the aquatic ecosystem of the Rio Tapajos
is from erosion of natural soils in the catchment rather than
from anthropogenic pollution. These results, in combination
with evidence that rates of erosion have been enhanced by
anthropogenic deforestation in the catchment (Roulet et al.,
1998, 1999), suggest that the input of natural mercury coming
from soils into the aquatic ecosystem may have increased over
historical levels in the region. This increase of total Hg in
aquatic ecosystem could potentially account for high concentrations of MeHg in fish.

74

H.do A. Kehrig et al. / Environmental Pollution 154 (2008) 68e76

The analysis of variance has shown significant differences

(N 42; H 13.11; p < 0.001 and H 12.99; p < 0.001)
in the normalized concentrations of total Hg and MeHg respectively (Fig. 1) but not for the percentage of MeHg to total
Hg (N 42; H 3.82; p > 0.10) across the four sampling
points of the Rio Tapajos. No significant differences were
found in the normalized concentrations of MeHg or the
percent MeHg to total Hg (N 24; Z 0.86; p > 0.10;
Z 1.29; p > 0.10, respectively) in Cichla spp. between the
samples collected at the vicinity of Itaituba and Jacareacanga
cities in the Rio Tapajos, where there are intense gold mining
activities. In contrast, significant differences were found in
normalized concentrations of MeHg between Cichla spp. collected near Itaituba and Santarem cities (N 16; Z 2.73;
p < 0.01) and near Jacareacanga and Santarem cities
(N 24; Z 3.03; p < 0.01) in Rio Tapajos, but no amongsite differences were found for the percent of MeHg to total
Hg ( p > 0.10).
Methylmercury concentrations in Cichla spp. decreased
(see Table 3) from the middle of Rio Tapajos at the vicinity
of Itaituba city to further downstream at Santarem city. Previous studies have found this same trend (Malm et al., 1995;
Uryu et al., 2001). It could be due to the possible influence
of the high mercury discharges in the middle of the Tapajos
River; intensive gold mining activity has been executed here
over the last 30 years and it is still considered a major gold
producing area in Brazil. The fluvial lacustrine system near
Santarem city can be considered an area without direct
influence of gold mining activities (Lima et al., 2000) and is
located downstream from the contaminated area in the middle
of Rio Tapajos.
The ManneWhitney test analysis has shown no significant
differences for the normalized concentration of MeHg
(N 18, Z 1.42, p > 0.10) and the percent MeHg to total
Hg (Z 1.24, p > 0.10) in Cichla spp. between the samples
collected at the vicinity of Santarem city and Lago Cury in
the Rio Tapajos. Lago Curi is a floodplain lake of the middle
Rio Tapajos, located close to Brasilia Legal city. It has been
subject to human disturbance for 100 years, being subject to
greater inputs of mercury, due to the ongoing disturbance of
soils in its catchments and the bottom sediments.
According to Howard (2001), dietary exposure to MeHg
was likely to be an important factor explaining the differences
in accumulation of methylmercury in Cichla. Probably, this is
because the sampling areas exhibit different sites for the
methylation of mercury and its incorporation into the food
chain. At Curi, the site was likely to be the aquatic macrophyte
roots, whereas at the forest sites it would be the submerged
litter layer. According to Guimar~aes et al. (2000), the floating
macrophyte roots were more important mercury methylation
sites, particularly those heavily colonized with periphyton
than the flocculent sediment settled over the litter layer.
3.4. Balbina and Tucuru reservoirs
Balbina reservoir, located about 200 km north-east from the
city of Manaus, was flooded in 1989. It is a relatively young

man-made reservoir and has intense fishing activity. Cichla

spp. are the predominant local fish species. The normalized
concentration of MeHg in the muscle of Cichla spp. collected
at Balbina hydro electrical reservoir, an area not impacted by
gold-mining, were similar to those found in the specimens
from the rivers impacted by human activities, Rio Tapajos
and Rio Madeira (Z 0.13, p > 0.10; Z 1.81, p > 0.05,
respectively) (Table 2). The Post hoc test analysis has shown
no significant differences for the normalized concentrations
of MeHg (Z 1.92, p > 0.05) in Cichla spp. between the
samples collected at the Balbina reservoir and Rio Negro. In
contrast, Kehrig and Malm (1999) showed that MeHg in
different piscivorous fish species, including Cichla spp., of
Balbina reservoir are among the highest in the Amazon.
This concurs with Boudou et al. (2005) as they observed up
to eight-fold higher mercury concentrations in fish caught in
a hydroelectric reservoir than in two rivers of Amazonian
French Guyana.
The Post hoc test analysis has shown a significant
difference (Z 3.28, p 0.001) in the normalized MeHg concentrations between Cichla spp. collected from the reservoirs
of Balbina and Tucurui. The Tucuru reservoir, located about
110 km northeast of Serra Pelada gold mining area, was
flooded in 1984 and Cichla spp. samples from there had higher
normalized MeHg concentrations than those from the Balbina
reservoir (Table 2). However, no significant differences
( p > 0.10) were observed between the percent MeHg to total
Hg found in muscle of Cichla spp. from these two reservoirs.
In addition, recent mercury data compared with those of
15 years ago from the Tucuru showed a reduction in mercury
concentrations by a factor of two or three times for piscivorous
fish, including Cichla spp. (Malm et al., 2004).
One of the parameters which may affect methylmercury
concentration in fish from Balbina and Tucuru is the age of
the reservoir. Newly-impounded reservoirs have high organic
matter content in water which increases methylation of
mercury by bacteria and results in higher bioaccumulation
and mercury concentrations in fish (Porvari, 1998; Schetagne
et al., 2000; Verdon et al., 1991). Shortly after inundation,
fish mercury concentrations clearly increased and remain
above background concentrations for 15e25 years (Porvari,
1998; Rolfhus et al., 2001) because soil and wetland inundation is an important source of MeHg to overlying water, as
well as to the aquatic food chain. The main expected mercury
sources to Tucuru and Balbina watersheds are the over
flooded soil leaching and atmospheric deposition.
4. Conclusions
In this study we showed that some biological characteristics
(body weight, standard length) affect concentrations of MeHg
in Cichla spp. muscle, whereas sex and different morphotypes
did not. We sampled three major tributaries of Rio Amazonas
(Tapajos, Madeira and Negro) and two reservoirs (Balbina and
Tucuru) within the Brazilian Amazon with different limnological conditions and levels of human activity (deforestation
for agricultural projects, damming for a hydroelectric power

H.do A. Kehrig et al. / Environmental Pollution 154 (2008) 68e76

plant, and alluvial gold extraction), and found significant

differences between the weight normalized MeHg concentrations in fish from the Tucuru and Balbina reservoirs and
also between the Tucuru reservoir and the three rivers. However, no differences were found among the weight normalized
MeHg concentrations in fish from the three rivers. Total Hg
determination is the most cost effective means of estimating
MeHg in fish, and is adequate for the purpose of assessing
patterns of accumulation in fishes.
Acknowledgements
The authors would like to thank the financial support of
Conselho Nacional de Desenvolvimento Cientfico e Tecnologico (CNPq). The authors thank two anonymous referees and
Karen Kidd for their help in improving an earlier version of
the paper.
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