Chemosphere 152 (2016) 328e334

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Chemosphere
journal homepage: www.elsevier.com/locate/chemosphere

Assessment of water quality and genotoxic impact by toxic metals in

Geophagus brasiliensis

ssio Resende Morais a, Stephan Maltano Carvalho b, *, Galber Rodrigues Araujo a,
Ca
Henrique Nazareth Souto c, Ana Maria Bonetti a, Sandra Morelli a,

rio Campos Jnior a
Edimar Olega
a
b
c

^ndia, Campus Umuarama, 38.900-402, Uberla

^ndia, Minas Gerais, Brazil
Institute of Genetics and Biochemistry, Federal University of Uberla
Department of Entomology, Federal University of Lavras, PO Box 3037, 37.200-000, Lavras, Minas Gerais, Brazil

rio Palm

Department of Cell Biology, Carmelitana Ma
erio Foundation, Jardim Zenith, 38.530-000, Monte Carmelo, Minas Gerais, Brazil

h i g h l i g h t s

The genotoxic potential was studied on Mumbuca river.

The Mumbuca stream is the target of different anthropic contaminants.

Toxic metals (Cr, Pb, Al and Ni) were identied and should be the responsible for DNA damage in Geophagus brasiliensis.

G. brasiliensis is an excellent indicator of environmental impacts in water bodies.

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 9 December 2015
Received in revised form
29 February 2016
Accepted 1 March 2016
Available online 15 March 2016

This study used the pearl cichlid Geophagus brasiliensis as a bioindicator to survey the health of the
aquatic environment on four sites (P1, P2, P3 and P4) of the Mumbuca stream located at Monte Carmelo/
MG, Brazil. The selection of different sites was made with reference to the gradient of urban activity and
via physicochemical and biological evaluation of water quality and genotoxicity. The water quality index
was classied as good for P1 and P4, regular in P2 and poor for P3. The micronuclei (MN) frequency
obtained from blood analysis was in agreement with the water quality, such that the higher values of MN
were detected in sites evaluated as poor. Water degradation conditions worsen according to the ow of
the stream over the sites P1, P2 and P3, but for site P4, located after the Monte Carmelo Sewage
Treatment Plant, improvements in the micronuclei frequency are detected. Our results showed high
levels of potentially toxic metals (chromium, lead, aluminum and nickel) in specic stream sites (P2 and
P3). We suggest that the micronuclei induction in G. brasiliensis could be due to the presence of these
compounds.
2016 Elsevier Ltd. All rights reserved.

Handling Editor: David Volz

Keywords:
Body water
Environmental contaminants
Water quality
Micronuclei
Toxic metals

1. Introduction
Among the existing natural water resources, 97% is saline, 2% is
present in reservoirs located on glaciers, and only 1% is composed
by fresh water present in groundwater, rivers and lakes. However,
the small percentage of accessible water bodies available for human
consumption is in an intense process of restriction due to pollution,
thus making these waters inappropriate for human and animal use

* Corresponding author. UFLA, Departamento de Entomologia Caixa, Postal 3037,

37.200-000, Lavras, MG, Brazil.
E-mail address: stephan.carvalho@den.ua.br (S.M. Carvalho).
http://dx.doi.org/10.1016/j.chemosphere.2016.03.001
0045-6535/ 2016 Elsevier Ltd. All rights reserved.

~o, 2002).
(Moraes and Jorda
Pollution of suchwater bod is largely occurring due to human
activities including the disposal of organic efuents from household wastewater (Camargo et al., 1995; UNCED, 1996) and industrial
efuents containing heavy metals and toxic compounds. Furthermore the disposal of efuents from agricultural activities (Arias
et al., 2007), play a role in indirectly contributing to the lotic
degradation process (Rashed, 2001; Marques et al., 2007). According to Rodrigues and Castilhos, 2003, since aquatic organisms
(e.g. sh) are part of the diet of man, transfer of pollutants (e.g.
heavy metals) via the food chain may be of great risks to humans,
potentially causing muscle and nerve damage.

C.R. Morais et al. / Chemosphere 152 (2016) 328e334

The Brazilian National Environment Council (CONAMA),

through the Resolution Number 357/2005, states that the water
bodies are subject to quality evaluation, employing physical and
chemical quantitative criteria which do not harm the organisms
that utilize this resource. These criteria are based on three different
aspects: (i) physical parameters (turgidity, color and temperature),
(ii) chemical parameters (pH) and (iii) biological parameters (Brasil,
2005; Boyd and Tucker, 1992; WHO, 2011).
In addition to traditional methodologies, ecotoxicological
studies are excellent alternatives to verify the quality of water and
~es
the effects that pollutants may cause on the ecosystem (Magalha
and Ferr~
ao Filho, 2008). Despite the deployment of Sewage Treatment Plant units, several pollutants (e.g. toxic metals and pesticides) are still dispersed in the water, requiring complementary and
specic procedures for monitoring the quality of water (Freire et al.,
2000). On the other hand, some techniques used for water treatment, may contribute to the formation of toxic compounds
(Tabatabai, 1986). In this context, the use of organisms (e.g. plants
and animals) as bioindicators becomes important to assess the real
risk of exposure to pollutants, especially since these organisms are
evaluated in their natural habitat (Buss et al., 2003; Nigro et al.,
2006). The tools to determine the levels of degradation of water
resources and the biota are key to ensuring the quality of this nite
resource.
In this way, sh are utilized as models for assessing the impact of
xenobiotics in water bodies. Additionally, they may be used in
toxicokinetics studies (distribution, metabolism and bioaccumulation) as well as in genotoxic, carcinogenic and teratogenic
assessment (Barsiene et al., 2015, 2014, 2012 and Scalon et al.,
2010). With increasing concerns about the genotoxicity caused by
pollutants, the use of sensitive bioassays can be a valuable tool in
assessing the genotoxicity occurrence of living organisms growing
in contaminated body waters. In this context, the micronucleus
assay has been widely used to investigate the genotoxic effects of
pollutants on DNA integrity (Gutierrez et al., 2015; Paulo et al.,
2015; Benameur et al., 2011 and Kapka et al., 2007).
According to Van Dyk et al. (2007); Wu and Deng (2006) and
Barsiene et al. (2015) aquatic organisms are excellent indicators of
genotoxicity, from a single drop of blood it is possible to visualize
thousands of sh erythrocytes. Thus, such methods provide excellent alternatives for the diagnosis of chromosomal aberrations or
even for micronuclei (MN) detection. The MN test is used to survey
the genotoxic impact of xenobiotics based on its damage on fragments of chromosomes, or entire chromosomes that occurs during
the cell division process. The micronuclei can be originated from
the failure of the mitotic spindle and development of nuclear
membranes containing DNA fragments that represents chromo~o et al., 2006). The MNs
somal breakage and damaged (Pantalea
originate after the organism is exposed to toxic agents present in
the ecosystem, so the micronucleus assay is widely used in biomonitoring practices, since it has high ability to detect cytogenetic
damage induced by genotoxic agents (Bolognesi et al., 2006;
~z et al., 1987; Ramirez et al., 1999).
Carvalho et al., 2002; Muno
The sh Geophagus brasiliensis Quoy and Gaimard, 1824 (Perciformes, Cichlidae), predominantly inhabit freshwater environments. They are appreciated as food, besides being important in
shing livelihoods (Beatty et al., 2013). These sh are extremely
sensitive to contaminants and for this reason can be potentially


used for water quality biomonitoring (Arantes et al., 2009; Beninca

rio et al., 2014; Voigt et al., 2015).
et al., 2012; Oso
Therefore, considering the interest to assess the impact of pollutants dispersed in water, this study has monitored the physical
and chemical quality of water from different collection sites in an
urban stream of Monte Carmelo, state of Minas Gerais, Brazil. The
genotoxic effect of the pollutants on G. brasiliensis was assessed by

329

means of the micronuclei approach.

2. Materials and methods
2.1. Sampling sites
Samples were collected at the city of Monte Carmelo, located in
the region of the Upper Paranaiba region (18 440 500 S and
47 290 4700 W), state of Minas Gerais, Brazil. The last demographic
census reported it has 45.772 inhabitants and a total area of
1.343.035 km2 (IBGE, 2010). The sites of collection for both water
and sh were dened according to existing human activities, being
classied as urban, industrial or agricultural activities. The location
of each site can be seen in Fig. 1.
Site 1 (P1): Located on the Sete Marias bridge, is a site that does
not undergo the process of anthropogenic degradation, as previously rated by the physicochemical analysis of the water and for
this reason was considered as a reference site (negative control) in
this study. Moreover, this site is also used by the Municipal
Department of Water and Wastewater for the catchment and distribution of water for local people (18 44019.700 S and 47 290 47.600
W).
Site 2 (P2): Located near the city downtown (18 430 36.700 S and

47 290 35.900 W) with continuous environmental impact due the
urban activities. Pollution at this site is the result of the domestic
sewage deposition in Mumbuca stream.
Site 3 (P3): Located after the city downtown and before the
wastewater treatment plant (18 420 86.300 S and 47 290 67.600 W).
This site is sewage target of domestic and industrial origin and
receives water from other water resources of the region (e.g. Santa
Maria stream).
Site 4 (P4): Located after the wastewater treatment plant, parallel to the MG-190 highway (18 41024.600 S and 47 280 98.100 W).
This site is a strong candidate to be contaminated by agricultural
residues due the proximity to several farms.
2.2. Fish sampling
A total of 60 pearl cichlid sh G. brasiliensis (30 males and 30
females/mean weight: 35 4 g and mean length: 15 5 cm) were
collected (12 individuals in each site of collection) at the wet season
(summer). The period of collection was intense drought, which
continued throughout the year.
Additionally, 12 individuals were sampled at the site P1, the site
that is reference without contamination, and were treated with
cyclophosphamide to be used as a positive control.
The sh collection was conducted using reel, line (0.30 mm),
hook and the annelid Pontoscolex corethrurus Muller, 1857 (Annelida, Glossoscolecidae) as bait. Samples were transported alive to

rio Palme

rio Founthe cytology laboratory of the Carmelitana Ma
dation, Monte Carmelo, MG, Brazil. At the laboratory, the individuals were kept at an aquarium (90  50  45 cm) with
permanent oxygen level control.
2.3. Physicochemical analysis of water
A total of 5 samples of water were collected in each site of
sampling. Physicochemical parameters of suspended solids and
biochemical oxygen demand (BOD) of water were analyzed according to the procedures reported by Boyd and Tucker (1992). The
evaluation of other physical and chemical parameters occurred
under the standard methods guidelines from Examination of Water
and Wastewater (APHA-AWWA-WPCF, 1998). Analyses of toxic
metals were made using a Varian Spectra AA-220FS atomic absorption spectrometry. Reference solutions of toxic metals were

330

C.R. Morais et al. / Chemosphere 152 (2016) 328e334

Fig. 1. Sites of water and biological material collection in the Mumbuca stream. P1, P2, P3 and P4 were the sites of sampling. STP is the Sewage Treatment Plant.

arranged in order of analysis, prepared with 0.9 mol/L of HNO3 and

CFA-C 10% v/v (pH 8.0) (limits of detection/LOD of 0.001 mg/L).
All analyzes were carried out at the Chemistry Laboratory of the
^ndia, MG, Brazil. The results of the
Federal University of Uberla
physicochemical analyses were compared with the tolerance limits
for each parameter according to Guideline 357/2005 of CONAMA
(CONAMA, 2005). These permissible limits are compatible with the
standards set by the World Health Organization (WHO, 2011).
2.4. Water quality index
Water quality index determination was carried out according to


the parameters of the Instituto Mineiro de Gest~
ao das Aguas
(IGAM,
2013). Data analyses were performed using the IQADATA Software,
available from the University of Santa Cruz do Sul, Rio Grande do
Sul state, Brazil. The water quality index is determined by the
product of quality established for each parameter such as: dissolved
oxygen, fecal coliform, pH, biochemical oxygen demand, nitrate,
phosphate, water temperature, turbidity and total solids (Yogendra
and Puttaiah, 2008). Based on this approach, the levels of water
quality are classied as: excellent (90 < IQA 100); good
(70 < IQA  90); poor (25 < IQA  50) and inappropriate (0 < IQA
25).
2.5. Genotoxicity analysis by micronucleus assay
For genotoxicity analysis, blood sample was collected from sh
tail vein with the aid of heparin syringe with an ultra-thin walled
needle. The material was xed to a slide, subjected to blood smear
technique and xed in ethanol for 10 min. Then, staining of cells
was carried out using 5% Giemsa in sodium phosphate buffer (pH
6.8) for 10 min, washed under running water, dried at room temperature and analyzed by optical microscopy at 1000x magnication (Francabandieira et al., 2007). The circular fragments

disconnected from the eukaryotic nucleus and that showed the

same patterns of color and intensity were considered as a micronuclei (Fenech et al., 2003). A total of 1000 cells per slide were
analyzed, totaling 12,000 cells per site of collection.
The 12 sh collected in P1 were used as a positive control,
receiving intraperitoneally the injection solution of cyclophosphamide at 0.04 mg/g of mass body. This treatment is responsible
for inducing genotoxic damage as described by Itoh et al., 2015, Nai
et al., 2015 and Dias et al., 2014. The erythrocytes were analyzed
72 h after drug administration.
2.6. Data analysis
To calculate the frequency of micronuclei occurrence among the
different samples, the data obtained were analyzed using the BioStat (AnalystSoft) software and the means were compared by the
parametric Tukey test at the signicant level of 0.05.
3. Results
3.1. Water physicochemical analysis and heavy metals detection
The results of the parameters analyzed for water quality in the
four sites of collection are presented in Table 1. In P1 (negative
control), where water is used for local supply, the physical and
chemical parameters were classied as class 1 (good quality). From
P2, intermediate parameters were conrmed at class 2 (regular
quality) for dissolved oxygen (5.15 1.28 mg/L), BOD
(4.8 2.15 mg/L) and fecal coliforms (NMP) (644 112 NMP/
100 ml).
In P3, values above to the established as standard for good water
quality (above the reference limits) were observed, highlighting to
the parameters fecal coliforms (1460 117 NMP/100 ml) (class 2),
dissolved oxygen (2.24 1.62 mg/L), BOD (13.0 2.16 mg/L) and

C.R. Morais et al. / Chemosphere 152 (2016) 328e334

331

Table 1
Physicochemical parameters and toxic metals contents in the Mumbuca stream water at sites P1, P2, P3 and P4 sites.
Sites of collection*

Parameters

P1
pH
Ammoniacal nitrogen (mg/L)
Surfactants (mg/L)
Dissolved oxygen (mg/L)
COD (mg/L)
BOD (mg/L)
Fecal coliforms (NMP/100 ml)
Nitrate (mg/L)
Total solids (mg/L)
Turbidity (UNT)
Temperature ( C)
Lead (mg/L)
Chromium (mg/L)
Aluminum (mg/L)
Nickel (mg/L)
Total phosphorus (mg/L)

7.03
0.23
0.07
7.12
17.45
1.2
126
0.03
35
2.98
22.17
0.009
0.01
0.10
0.006
0.01

MLL
P2

1.65 a
0.07 a
0.03 a
1.08 a
6.61 a
1.03 a
101 a
0.01 a
17 a
1.44 a
1.52
0.001 a
0.00 a
0.06 a
0.003 a
0.00 a

6.31
0.25
0.08
5.15
24.43
4.8
644
0.11
76
14.95
22.04
0.011
0.02
0.20
0.008
0.02

P3

1.65 a
0.14 a
0.02 a
1.28 b
5.25 a
2.15 b
112 b
0.04 a
15 a
3.12 a
4.53
0.006 b
0.00 a
0.09 b
0.001 a
0.01 a

6.04
1.08
0.39
2.24
27.25
13.0
1460
0.57
158
48.65
23.08
0.022
0.06
0.45
0.048
0.6

P4

1.02 a
0.63 a
0.15 a
1.62 c
6.07 a
2.16 c
117 c
0.20 a
24 a
7.25 b
1.66
0.008 b
0,03 c
0.11 c
0.022 c
0.01 c

6.59 1.12 a
0.07 0.03 a
0.04 0.01 a
7.01 2.07 a
12.28 8.13 a
1.7 1.12 a
94 68 a
0.01 0.03 a
11 10 a
15.62 8.35 a
23.65 2.43
< LOD
< LOD
0.10 0.05 a
0.001 0.000 a
0.001 0.000 a

6e9
3.7
0.5
Not inferior to 6
Not inferior to 6
3
200
10
500
40
e
0.01
0.05
0.1
0.025
0.1

MLL: Maximus limits stipulated by the Legislation, BOD: Biochemical Oxygen Demand, COD: Chemical Oxygen.
Demand, < LOD: above limit of detection (0.001 mg/L).
*
Means Standard Deviation followed by the same letter are classied according to the index proposed by the directive CONAMA (2005): a class 1 (good quality), b class 2
(regular quality) and c class 3 (bad quality).

total phosphorus (0.6 0.01 mg/L) belonging to the class 3. In this

site, the values obtained of dissolved oxygen; DBO and total
phosphorus, were crucial to the impairment of water quality. Parameters of class 2 for turbidity (48.65 7.25 UNT) were observed
in this site.
In P4, values above the standards established for good water
quality for all of the physicochemical parameters analyzed were not
observed (Table 1). This site was referred as belonging to class 1.
The presence of potentially toxic metals was mainly concentrated at P3, with high levels of chromium (0.06 0.03 mg/L) (class
3), aluminum (0.45 0.11 mg/L) (class 3), nickel (0.048 0.022 mg/
L) (class 3) and lead (0.022 0.008 mg/L) (class 2) (Table 1). In P2
site were found lead (0.011 0.006 mg/L) and aluminum at level of
0.20 0.09 mg/L (class 2) (Table 1).
Table 2 summarizes the index of water quality of Mumbuca
stream, taking as reference the IGAM (2013) parameters. The value
of the water quality was considered poor (41.39) in P3. In P1, P2 and
P4 sites, the water quality was classied as good (80.05), regular
(62.62) and good (78.25), respectively.
3.2. Genotoxicity analysis
The frequency of MN detected in G. brasiliensis collected from
the Mumbuca stream is presented in the Table 3. The frequency of
MN was signicantly higher (P < 0.05) in the positive control (PC)
(19.33 3.72), P3 (11.50 2.23) and P4 (9.41 2.27). In contrast, in
sample from sh collected from P1 site that was used as negative
control, the frequency of MN was lower (0.83 1.02) compared to
Table 2
Water Quality Index obtained in the Mumbuca stream analysis based on the pa

rameters established by the Instituto Mineiro de Gest~
ao das Aguas
(IGAM).
Site of collection

Geographic coordinate

Quality values

Classication

P1

18 440 19.700 S

47 290 47.600 W
18 430 36.700 S
47 290 35.900 W
18 420 86.300 S
47 290 67.600 W
18 410 24.600 S
47 280 98.100 W

80.05

Good

62.62

Regular

41.39

Poor

78.25

Good

P2
P3
P4

sample from sh collected in other sites. Additionally, there was no

signicant difference (P > 0.05) between the frequency of MN
detected in sh from the sites P1 and P2 (3.33 0.77) (Table 3).
4. Discussion
Among several pollutants, toxic metals have a special importance due to the fact that this group of minerals is very harmful to
humans, plants and animals. According to Roche and Boge (1996),
toxic metal contamination in lotic environments can affect and
have detrimental effect on the ecology. Various studies have shown
genotoxic effects in aquatic organisms exposed to contaminated
water with toxic metals (Yadav and Trivedi, 2009; Barbosa et al.,
2009 and Matsumoto et al., 2006). In the present study, toxic
metals were found in sites of the Mumbuca stream, which suffers
with the impact of anthropogenic activities.
The physicochemical characteristics of the water analyzed in
this study presented changes in various parameters, showing the
direct inuence of human activities in the sites analyzed. In P1, the
water quality was classied as good, since there was no apparent
process of environmental degradation, and in this investigation this
site was considered as reference point (negative control). The
clastogenic and/or aneugenic effects identied by the MN test
support the results obtained from water analysis in P1, since the
chemical parameters analyzed did not present high concentrations
of potentially toxic metals and/or other xenobiotic substances that
could lead to DNA damage. The P2 is located on a stretch of the
Mumbuca stream that suffers with the deposition of sewage and
household waste. The rate of fecal coliform in P2, as well as in P3,
was the highest and these sites were classied as Class 2 and 3,
respectively. According to the water quality index, these sites presented regular and poor environmental quality, respectively.
When compared to the negative control (P1), P2 did not show
signicant differences for the MN frequency in G. brasiliensis, which
is consistent with the results obtained for physicochemical and
toxic metals analysis. Although not different from control, the
occurrence of MN at P2 may have been induced by the presence of
lead and aluminum (class 2), which are compounds recognized for
their ability to induce DNA damage (Skerfving, 1993). The ceramics
industry, has been identied as a potential route for lead deposition
(Paoliello and Capitani, 2003), hence with a strong industrial

332

C.R. Morais et al. / Chemosphere 152 (2016) 328e334

Table 3
Micronucleus (MN) frequency in erythrocytes of Geophagus brasiliensis collected in the different sites along the Mumbuca stream.

Site of collection

Number of individuals

Total cells

MN frequency % SD*

P1
P2
P3
P4
Positive control

12

12,000

1.02
03.33
11.50
09.41
19.33

0.83
0.77
2.23
2.27
3.72

a
a
b
b
c

Total MN
10
40
138
113
232

Means Standard Deviation followed by the same letter in column do not differ by the parametric Tukey test (P  0.05).

presence in Monte Carmelo city, ceramics may be indicative of the

pollution in this area. In addition to being a potential genotoxic
agent, studies with animal models have reported lead to be a
carcinogen, with exposures resulting in kidney and brain tumors
(Skerfving, 1993). The presence of lead in P2 and P3 observed in this
study may be responsible for DNA damage, as was already reported
by Alves et al. (2006) in wolf sh Hoplias malabaricus Bloch, 1794
(Caraciformes, Erythrinidae).
In P2 and P3, aluminum (Al3) levels were above the tolerable
values as determined by CONAMA (2005). According to the CONAMA Resolution number 357/2005, for Al3, water categorized into
class 3 is t for human consumption due to prior treatment via a
conventional or advanced system. Toxic effect on the central nervous system occurs with the ingestion of water with Al3 at high
rates (CETESB, 2006). Being located after central area of Monte
Carmelo, the P3 site showed higher irregularity with regard to
parameters established by CONAMA, since all pollutants derived
from domestic and industrial residues accumulate in this site.
The frequency of MN detected in sh collected from P3 was
higher when compared to the negative control. The frequency of
MNs detected at P3 corresponds with the chemical analysis of the
water, showing the presence of potentially toxic metals to be above
the permissible limits. In parallel, this site was warranted with poor
water quality index. Various genotoxic agents such as chromium,
lead, aluminum and nickel, were found at this site, indicating that
these compounds are responsible for the induction of MNs in sh
erythrocytes. High levels (class 3) of chromium were found at this
site. Taking in account what has already been reported (Russo et al.,
2004), we can hypothesize that the increased DNA damage rate in
erythrocytes found in sh collected at this site might be due the
high concentrations of chromium. The increased nuclear abnormalities have also been detected in sh after exposure to signicant
amounts of chromium (Matsumoto et al., 2006). Chromium has
genotoxic potential (Maeng et al., 2004), can alter the human immune system (Shrivastava et al., 2002) and induce throat cancer;
nasal lesions and perforation of the septum and skin; kidney and
liver disorders. This compound can be fatal by causing disturbance
in the gastrointestinal tract of humans exposed to such chemical
element (ASTDR, 2002). Several studies have shown that exposure
to chromium can result in DNA damage (Von Burg and Liu, 1993;
Stohs and Bagchi, 1995; Mount and Hockett, 2000).
According to Rank and Nielsen (1998), nickel is a genotoxic
agent and induces the formation of MNs in aquatic organisms
(Okunola et al., 2015). This metal was identied at levels above of
the acceptable limits in the P3 site, and can be a factor to the high
frequency of MNs detected. It is reasonable to say that at P3 there
are elevated hazard regarding genotoxic effects more than is
observed in the other analyzed sites and that the main factor of
these disturbances is the presence of toxic metals in the water.
According to Barbosa et al. (2009) and IARC (1996) toxic metals are
able to damage the genetic material of exposed organisms.
Toxic metals are environmental pollutant that can harm the
health of sh. Several studies have demonstrated the bioaccumulation capacity of these pollutants on aquatic organisms

(e.g. sh). For example, these compounds may lead to histopathological changes (gills, liver and kidney), increase on oxidative stress,
damage in the DNA and associations with many sh deformities in
natural populations causing devastating effects on survival, growth
rates and welfare (Fonseca et al., 2016; Javed et al., 2016; Abalaka,
2015; Nunes et al., 2015; Sfakianakis et al., 2015). According the
survey of Campos Jnior et al., 2015, was not detected the pesticide
residues on Mumbuca stream along the urban area, suggesting that
samples of sh that showed DNA damage might be correlated with
the presence of toxic metals.
Dhanakumar et al. (2015) assessed the toxic metal bioaccumulation in six commercial sh species. In this study, two
species in particular presented the highest accumulation of toxic
metals. If included in the diet of humans, contaminated sh offer
high risk to human health (Zhu et al., 2015).
In this work, we found that the degradation of the aquatic
environment induced by human activities intensies along the
course of the stream, from P1 to P3. This is shown by the sustained
decrease in values of water quality index combined with different
quality conditions at each site evaluated. There is an improvement
in the parameters of water quality and, consequently, better quality
of water in P4, a site underlying the Sewage Treatment Plant. According to the chemical analysis of the water and the water quality
index (good quality) for the P4 site, that is located after the Sewage
Treatment Plant, it indicates that the wastewater treatment plant is
being effective in reducing water pollution. However the MNs
frequency detected in sh collected at this site was statistically
different when compared to the negative control. We hypothesize
that this difference may be related with another kind of xenobiotics
capable to induce DNA damage. The presence of pesticides which
were not analyzed in this study can also be responsible for inducing
a MN frequency above the threshold tolerated by sh. It has already
been reported (Matsumoto et al., 2006; Shaw and Handy, 2006; Atli
and Canli, 2007) that DNA damage occurs when animals are
exposed to environments containing genotoxic compounds or even
by ingestion of others contaminated aquatic animals. This hypothesis is supported also by Stefani et al. (2008), who claim that
the diet of G. brasilienses consists mainly of vegetables, aquatic invertebrates and larvae of other sh. Furthermore, Buss et al. (2003)
reports that even with parameters within the standards established
by CONAMA, some of these parameters may interfere with the
biological relationships and can generate problems for living
organisms.
In this work we have not evaluated directly the presence of
pesticide residues in the water. However, due the agricultural activity at this city (mainly on coffee farming) and the proximity to
the urban area, pesticide residues may be responsible for the induction of MNs in sh erythrocytes. Several studies conrm
mutagenic and genotoxic effects of pesticides in vivo and in vitro
(Tripathy et al., 1987; akir and Sarikaya, 2005; avas and Konen,
2007; Ghisi et al., 2011; Mishra et al., 2014; elik et al., 2014).
The results presented here demonstrate the intensication of
degradation of water body along the Mumbuca stream in Monte
Carmelo city. Evidence of potentially toxic metals and other

C.R. Morais et al. / Chemosphere 152 (2016) 328e334

changes in water quality along the stream indicate hazard for human consumption, animal and the environment. If we consider that
domestic, industrial and agricultural waste water do not pass
through the wastewater treatment plant, the contamination source
should be investigated and controlled.
In conclusion, the waste disposals arising from human activities
such as household waste, industrial and agricultural are factors that
are directly related to the process of degradation of water bodies
and biota. The identication of toxic metals via chemical analysis is
potentially responsible for the MN induction in G. brasiliensis.
Furthermore, due to its sensitivity to pollutants, this species of sh
can be used as an excellent bioindicator in programs aiming to
monitor the impact of anthropogenic activity on body waters.
Acknowledgements
~o de Amparo a
 PesThe authors would like thank the Fundaa
~o de
quisa do Estado de Minas Gerais (FAPEMIG), Coordenaa
Aperfeioamento de Pessoal de Nvel Superior (CAPES), Conselho

gico (CNPq),
Nacional de Desenvolvimento Cientco e Tecnolo
~o Carmelitana Ma

rio Palme

rio (FUCAMP) and the UniFundaa
^ndia (UFU) for providing operational
versidade Federal de Uberla
and/or nancial support to develop the research.
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