Ecotoxicology and Environmental Safety 253 (2023) 114670

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Ecotoxicology and Environmental Safety

journal homepage: www.elsevier.com/locate/ecoenv

Genotoxicity effects in freshwater fish species associated with gold mining

activities in tropical aquatic ecosystems
Ángel Cruz-Esquivel a, Sergi Díez b, *, José Luis Marrugo-Negrete a, *
a ́ Colombia
University of Córdoba, Carrera 6 No. 77–305, Monteria,
b
Environmental Chemistry Department, Institute of Environmental Assessment and Water Research, IDAEA-CSIC, E-08034 Barcelona, Spain

A R T I C L E I N F O A B S T R A C T

Keywords: The main aim of this study was to investigate total mercury (THg), methylmercury (MeHg) and arsenic (As)
Mercury concentrations, and their genotoxic effects on fish species in freshwater habitats impacted by gold mining ac­
Arsenic tivities in the Mojana and Bajo Cauca regions (Northern Colombia). A total of 255 individuals of Prochilodus
Micronucleus
magdalenae (PM) and Hoplias malabaricus (HM) were collected in different areas of northern Colombia, 205 in the
Erythrocytic nuclear alterations
Colombia
exposed groups: Mojana 1 (61), Mojana 2 (81) and Bajo Cauca (63); and 50 individuals in the control group.
Dorsal muscle was analysed for pollutants and blood to perform micronucleus (MN) and erythrocytic nuclear
alterations (ENA) tests. The results of the MN revealed statistically significant (p < 0.05) genetic damage in both
PM (Mojana 1 = 29.7 ± 14.2; Mojana 2 = 25 ± 6.25; Bajo Cauca= 26.6 ± 10.6) and in HM (Mojana 1 = 17.7 ±
7.8; Mojana 2 = 20.4 ± 6.3; Bajo Cauca= 20.8 ± 9.8) compared to the control group (PM= 10.5 ± 3.6; HM= 9.1
± 3.9). Likewise, the frequency of ENA was statistically higher in the exposed groups compared to the control
group (p < 0.05). On the other hand, the concentrations of THg, MeHg and As found in tissue samples were
significantly higher (p < 0.05) compared to the control group, being the Bajo Cauca region the area of highest
risk due to high concentrations of THg (651.2 ± 344.5 μg/kg for HM and 678.5 ± 983.9 μg/kg for PM) and MeHg
(504.6 ± 220.9 μg/kg for HM and 606.8 ± 886.4 μg/kg for PM). Results showed that mean THg values for both
species in Bajo Cauca exceeded the WHO maximum limit (set in 500 μg Hg/kg) in fish for human consumption.
Results suggest that DNA damage in erythrocytes is associated with the presence of Hg, MeHg and As, coming
from mining activities.

1. Introduction adverse health effects on organisms such as oxidative stress, endocrine

Pollution by toxic elements in aquatic ecosystems is a global serious
problem due to effects on human health and wildlife (Barone et al.,
2018). Mercury (Hg) and arsenic (As) are considered toxic trace ele­
ments that can reach aquatic ecosystems from natural or anthropogenic
sources such as agricultural and mining activities, causing adverse ef­
fects because of their high toxicity (Córdoba-Tovar et al., 2022). In the
aquatic environment, Hg is easily transformed into its most toxic organic
form, methylmercury (MeHg), which has a high potential for bio­
accumulation in organisms and biomagnification in the food chain (Yan
et al., 2019). Therefore, pollutants as Hg, MeHg and As can have
extreme toxicity even at low concentrations leading to sublethal effects
or deaths in native fish populations (Jasim et al., 2016). Some elements
are essential for fish metabolism, while others such as Hg and As have no
known function in the biological system (Khan et al., 2016). In contrast,
disruption, DNA damage and cell death have been documented (Chen
et al., 2018).
Fish are an important food resource for human populations; how­
ever, in gold mining areas, their habitat receives high polluted dis­
charges from upstream activities (de Souza Neto et al., 2020;
Palacios-Torres et al., 2018). Unfortunately, toxic elements coming from
anthropogenic activities can be detected in several matrices (Díez et al.,
2011; Marrugo-Negrete et al., 2015, 2017) and bioaccumulate in fish
tissues (Marrugo-Negrete et al., 2008, 2018, 2020; Córdoba-Tovar et al.,
2022;). In tropical freshwaters, Hg and MeHg can be bioaccumulated in
some of the most popular fish such as Prochilodus magdalenae (boca­
chico) and Hoplias malabaricus (moncholo) (Marrugo-Negrete et al.,
2008, 2018). Additionally, As has also been found to bioaccumulate in
fish (Perera et al., 2016). This process generates impact at the cellular
level (e.g. oxidative stress and reactive oxygen species, ROS), which

* Corresponding authors.
E-mail addresses: sergi.diez@idaea.csic.es (S. Díez), jmarrugo@correo.unicordoba.edu.co (J.L. Marrugo-Negrete).

https://doi.org/10.1016/j.ecoenv.2023.114670

Available online 27 February 2023

0147-6513/© 2023 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-
nc-nd/4.0/).
Á. Cruz-Esquivel et al.
cause DNA damage and cell death (García-Medina et al., 2017).
P. magdalenae and H. malabaricus are of high economic, gastronomic,
cultural and commercial importance in Colombia, being the former an
endemic freshwater fish that occurs in the Magdalena, Sinú and Atrato
river basins (Mojica et al., 2002; Maldonado-Ocampo et al., 2005).
Moreover, P. magdalenae is an important fishing resource for artisanal
fishing communities, being one of the most commonly caught species.
This is detritivore species that feeds on fine bottom sediment containing
organic particles. It is considered a key element in the ecosystem and can
play an ecological role due to its sediment processing habit (Jar­
amillo-Villa and Jiménez-Segura, 2008). There are signs of population
reduction in catches and sizes of this reophilic fish, being considered a
vulnerable species (Mojica et al., 2012). On the other hand,
H. malabaricus is a freshwater neotropical fish that is widespread in
Colombia, inhabiting marshes, ponds, reservoirs and streams. It is an
interesting model for investigating sensitivity to toxic elements as a
predator that can influence fish communities from the top to the bottom
(Monteiro et al., 2013). This species is known for its resistance to hyp­
oxia, food deprivation and xenobiotic exposure (Monteiro et al., 2013;
Mela et al., 2007).
Fulton’s condition factor is widely used in environmental monitoring
and biology fish studies, to show growth rate and changes in nutritional
condition of fish. This factor is calculated from the relationship between
the weight of a fish and its length with the intention to indicate changes
in fish condition in response to contaminants and/or stress (Nahs et al.,
2006; Hina et al., 2021).
The genotoxic potential of metals can be evaluated by micronucleus
(MN) assay and erythrocytic nuclear alterations (ENA), which are
widely used as biomarkers in toxicological testing because they can
provide estimates of genetic damage, cell division errors, and cell death
(Abdel-Khalek, 2015; Anbumani and Mohankumar, 2019). The MN test
is the most widely used because it detects the genotoxicity of a wide
range of pollutants, especially in fish (Gomes et al., 2015; D’Agostini and
La Maestra, 2021), whereas the use of ENA has not been as extensively
studied as micronucleous (Ç avas and Ergene-Go¨zu¨kara, 2005). In this
sense, genetic damage is an indicator of the toxicity of cells in response
to stress by hazardous elements, so it is a molecular marker of the effects
of environmental contaminants. Specifically, MN in erythrocytes are
biomarkers of sublethal exposure to various xenobiotics, including metal
(loid) elements in aquatic organisms (Anbumani and Mohankumar,
2019), and micronucleous assay is considered the standard method for
assessing these effects in fish blood (Obe et al., 2002; Anbumani and
Mohankumar, 2019).
In aquatic ecosystems, fish are frequently used as indicator for
environmental monitoring, genotoxicity, and water quality studies, due
to their role in the food chain, their importance in human nutrition, and
their ability to biomagnify metals (Rashed, 2001; Lamas et al., 2007; Ali
and Khan, 2019; Córdoba-Tovar et al., 2022). Because freshwaters
ecosystems of Colombia are being affected by Hg and As contamination
(Olivero-Verbel et al., 2015; Marrugo-Negrete et al., 2018a), such kind
of studies are needed.
The main objective of this study is to assess, for the first time, the
impact of THg, MeHg and As in the Mojana and Bajo Cauca regions by
examining the incidence of micronucleous nuclear alterations in circu­
lating erythrocytes on two relevant fish species, P. magdalenae and
H. malabaricus. These species were selected because they are the most
abundant, caught and consumed fish species in riverine habitants in the
Mojana and Bajo Cauca regions.
2. Materials and methods
2.1. Study area
This research was carried out in two geographically extensive re­
gions of the Colombian territory. The Mojana region located in north­
western Colombia has an approximate area of 5545 km2
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Ecotoxicology and Environmental Safety 253 (2023) 114670
(Marrugo-Negrete et al., 2015). This region has a tropical wet and dry
climate, with temperatures ranging from 28◦ to 33◦ C. The region is
delimited by three rivers, the Magdalena, the Cauca and the San Jorge,
and it is crossed by streams that connect muddy complexes that receive
river waters during flood periods (Pinedo-Hernández et al., 2015). There
are no gold mining activities in the Mojana region. However, gold
mining waste is transported from the surrounding mining areas by the
streams or water flows that occur during the rainy season (Marrugo-­
Negrete et al., 2010, 2017, 2019).
For this study, the Mojana region was divided into two areas due to
its large size and hydrogeographic differences in each sub-region. A first
group called Mojana 1 is composed of the following municipalities: San
Marcos, San Benito de Abad, Ayapel, Caimito and Sucre. The main water
resources from fish samples were caught were the Ayapel swamp, the
San Jorge River, the Caimito swamp and the Sucre Sucre and San Benito
Abad swampy complex. A second group named Mojana 2 is formed by
the following municipalities: Guaranda, Achí and San Jacinto del Cauca.
The aquatic ecosystem from which the fish samples were obtained was
the Cauca River and surrounding marshes (Fig. 1). A third study area
was the Bajo Cauca of Antioquia. This is located at the northeastern end
of the Antioquia department. It is one of the nine sub-regions into which
Antioquia is divided, formed by the municipalities of Caucasia, Cáceres,
El Bagre, Nechí , Tarazá and Zaragoza. The territories covered by its
municipalities add up to 8485 km2 which corresponds to 13.5% of the
departmental territory. Gold mining, which uses Hg in the amalgam­
ation process for the recovery of the precious metal, has become the
main economic activity of the region, but it does not represent a sig­
nificant source of development, due to its high level of informality and
the social, economic and environmental problems that its exploitation
has brought with it (Agudelo-Echavarría et al., 2020). This has led to
countless impacts such as soil erosion and degradation, water resources
pollution, fauna and flora disappearance, and atmospheric pollution,
deforestation, flooding, among others (Cordy et al., 2011;
Marrugo-Negrete et al., 2019). The main water resources, where the fish
samples were obtained, were the Nechí River, the Cauca River and the
Tiguí River (Fig. 1). The control samples were caught in a swamp
ecosystem located in the lower Sinú River basin, with no gold mining
activities (Marrugo-Negrete et al., 2017).
Previous studies in the Mojana region for fish and sediments have
shown that around 12% (Marrugo-Negrete et al., 2018b), and 14%
(Marrugo Negrete et al., 2008) of the total fish samples exceeded the
permissible limit established by the World Health Organization (i.e.
500 μg Hg/kg), although recent studies (Romero-Suárez et al., 2022)
showed that none of the studied fish had Hg amounts above the estab­
lished limits. On the other hand, the mercury in the sediments of the
Mojana region exists primarily as the fraction associated with organic
mercury (Pinedo-Hernández et al., 2015), showing a contamination
degree from non-polluted to moderate pollution (Marrugo-Negrete
et al., 2018b).
2.2. Fish sample collection
The sampling period was from March to November 2018. All sites,
where fish samples were collected, were georeferenced (Fig. 1). The fish
use in this research was conducted in agreement with the national ani­
mal research ethics guide. A total of 255 individuals were collected, 205
in the exposed group in different areas of northern Colombia: Mojana 1
(N = 61), Mojana 2 (N = 81) and El Bajo Cauca (N = 63); and 50 in­
dividuals in the control group. Collections were carried out during a
transitory period with the help of local fishermen. After collection, the
fish length and total weight were recorded. Blood samples were obtained
by caudal vein puncture using a heparinized syringe. Blood was smeared
immediately onto two clean glass slides, air dried, and then fixed in
absolute ethanol for 20 min or in absolute methanol for 10 min (Porto
et al., 2005). The fish were packed, labeled and transported refrigerated
to the laboratory, where a sample of dorsal tissue was removed using
Á. Cruz-Esquivel et al.
Fig. 1. Study area.
sterile blades. The samples were lyophilized, homogenized by macera­
tion and stored in sealed bags at − 20 ◦ C until analysis (Mechora et al.,
2014).
2.3. Metals analysis in dorsal fish tissue
For total mercury (THg) analysis, 20–50 mg of dry and homogenized
sample were weighed, determining the concentration by using a DMA 80
TriCell Milestone Direct Mercury Analyzer using EPA method 7473.
Analysis of As was performed following a modification of the method
described by Szkoda et al. (2006), using sample calcination (1 g of
sample and Mg(NO3)2 as calcining reagent for As) at 550 ◦ C after
freeze-drying and maceration. Later the residue was digested until
dryness with HNO3 and re dissolving with HCl, to finally be quantified
by Thermo Scientific atomic absorption spectrometer model iCE 3500
with hydride generator (VP100) (APHA, 1998). For the MeHg analysis in
fish about 0.5 g sample was weighed in a centrifuge tube and mixed with
HBr and toluene. The organic phase was shaken for 2 min and separated
by centrifugation. Methylmercury was measured using a DMA-80
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Ecotoxicology and Environmental Safety 253 (2023) 114670
analyser according to a modification of EPA Method 7473 as stated by
Maggi et al. (2009). Concentrations of THg, MeHg and As were
expressed in µg/kg wet weight (ww).
For quality control methods 10% of the samples were analyzed in
duplicate and quantified using a calibration curve with a determination
coefficient greater than or equal to 0.9995; the detection limit was
determined as three times the standard deviation of 10 measurements of
a reagent blank, for As was 5 µg/kg, THg 0.13 µg/kg and MeHg 0.25 µg/
kg, respectively. In order to verify the traceability of the method it was
evaluated by analysing, in triplicate, certified reference material Dogfish
Muscle DORM-4 of the National Research Council (As: certified value =
6.80 ± 0.64 μg/g, found = 6.56 ± 0.23 μg/g; THg: certified value =
0.410 ± 0.055 μg/g, found = 0.402 ± 0.021 μg/g; MeHg: certified =
0.354 ± 0.031 μg/g, found: 0.372 ± 0.025 μg/g). The recovery per­
centages in DORM-4 were 95% for As, 98% for THg and 105% for MeHg,
with coefficients of variation lower than 5%, within the 95% confidence
limits.
Á. Cruz-Esquivel et al.
2.4. Micronucleus and ENA tests
The micronucleus, nuclear bud and binucleated determination was
performed with slight modifications according to the method described
by Fenech (2000). The samples were stained with 10% of Giemsa so­
lution for 10 min, air dried and permanently fixed. A total of 2000 red
blood cells were examined for each specimen under a light microscope
(OlympusBX43) with immersion oil at 1000 magnification. A total of
2500 erythrocyte cells per individual were counted, distributed in 5
slides (replicates) that served as quality control (500 x slide) to stan­
dardize the frequency micronucleus erythrocytes. Slides were scored by
a single observer using blind review. For MN scoring purpose, only
nonrefractive small nuclei (>1/3 of the main nucleus) located close to
the main oval nucleus of round erythrocytes with intact cytoplasm were
considered. Other types of ENA such as cells were also counted in a
similar way.
2.5. Statistical analysis
The normality of the results was evaluated using the Kolmogorov-
Smirnov test. The statistical analysis of the differences in THg, MeHg
and As concentrations among groups or collection area was performed
using the non-parametric Kruskall Wallis test with Dunn’s correction for
multiple comparisons. The condition factor in individuals was deter­
mined with the following equation: K = 100 W/L3; where W is the
weight in grams and L is the total length of the fish in centimeters
(Seiyaboh et al., 2016). A Spearman rank correlation analysis was per­
formed to establish an association between element concentrations in
P. magdalenae and H. malabaricus with weigth, length, condition factor
and genotoxic variables. Spearman’s test was performed to evaluate the
correlation between THg, MeHg, As, genotoxic variables and condition
factor. For all analyses, the established significance criterion was
p < 0.05. The analysis was done with Minitab version 16 statistical
software and StatGraphics Centurion XV Version 2.15.06.
3. Results
Table 1 show the mean values of the morphometric variables (length,
weight and condition factor) for H.malabaricus and P. magdalenae
collected in the three sites and control. Significant correlation was found
between length and weight for both species (p < 0.01). Carnivorous
H. malabaricus showed low values of condition factor (K), whereas
P. magdalenae samples have K values higher than 1.
Concentration values in the two species studied are presented in
Fig. 2. Results in the fish dorsal muscle reveal that THg, MeHg and As, in
aquatic ecosystems are influenced by mining activities, and were sta­
tistically higher to values of fish species at the control site (p < 0.05).
THg and MeHg concentrations in both P. magdalenae and H. malabaricus
were statistically higher in the Bajo Cauca region compared to Mojana 1
and Mojana 2 (p < 0.05). On the other hand, As concentrations found in
H. malabaricus were statistically higher in the Mojana 1 and Mojana 2
regions compared to the Bajo Cauca region (p < 0.05). In contrast, As
concentrations in P. magdalenae were statistically higher in the Mojana 2
region compared to the other regions.
The median values of MeHg concentrations found in muscle tissue of
H. malabaricus and P. magdalenae in Bajo Cauca exceeded the WHO limit.
Table 1
Morphometric variables for H. malabaricus and P. magdalenae collected in the three sites and control. Data are represented as means of eight samples ± SE.
Sample sites n H. malabaricus
Total length (cm) Weight (g)
Mojana 1 61 27.4 ± 3.2 191.5 ± 34.9
Mojana 2 81 29.1 ± 4.2 205.8 ± 48.6
Bajo Cauca 63 29.3 ± 1.5 227.2 ± 10.5
Control 50 28.3 ± 2.0 224.6 ± 18.1
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Ecotoxicology and Environmental Safety 253 (2023) 114670
Likewise, the median value MeHg found in H. malabaricus in Mojana 2
was also higher than the WHO limit to protect vulnerable people,
including pregnant women and children under 15 years old, established
in 200 μg/kg (WHO, 1990). The mean MeHg as a percentage of THg for
all the sites was 81.9 ± 9.5% (range 56.0–99.4%), indicating that
organic mercury was the predominant form of Hg in the muscle tissue of
fish.
A comparison between sampled sites in aquatic systems indicates the
following order from highest to lowest value in relation to THg and
MeHg concentrations in both species of fish: Bajo Cauca > Mojana
2 > Mojana 1. In general, lower concentrations were found in
P. magdalenae (Table 1), which may be related to their dietary habit (i.e.
is a detrivorous species that feeds on the fine bottom sediment); while
higher concentrations were found in H. malabaricus (carnivorous feeding
habit).
Micronucleous and the two nuclear alterations (i.e. nuclear buds and
binucleated) were identified in fish erythrocytes (Table 2). The highest
average values of ENA frequencies were observed in samples of fish of
the P. magdalenae species. In both species, statistically significant dif­
ferences in genotoxic variables were observed compared to the control
group. Therefore, all ENA were statistically higher in individuals
inhabiting water bodies impacted by mining activities compared to the
control group (p < 0.05). On the specie P. magdalenae, the frequency of
cells in micronuclei showed higher median values in the Mojana 1,
followed by Bajo Cauca region and Mojana 2. Furthermore, the fre­
quency of cells in nuclear buds showed the following decreasing order:
Bajo Cauca > Mojana 2 > Mojana 1. On the other hand, the frequency of
binucleated cells was in the following order: Bajo Cauca > Mojana
1 > Mojana 2.
For H. malabaricus, the region with the highest average value of MN
was Bajo Cauca, followed by Mojana 2 and Mojana 1. As for the other
ENA, the region that reached a higher average value was Bajo Cauca,
followed by Mojana 2 and Mojana 1.
Table 3 shows the correlations results between THg, MeHg and As
concentrations in muscle tissue with the length (L), weight (W) and
condition factor (K) of both fish species.
Positive and significant correlations were found between W and L of
the ictic species with THg and MeHg concentrations in muscle tissue.
However, for P. magdalenae, As concentrations show a negative associ­
ation with the W of the individuals (p < 0.05). Negative significant
(p < 0.05) correlations were recorded between condition factor K, with
THg, MeHg and As concentrations, in P. magdalenae. For H. malabaricus
these correlations were only significant for THg and MeHg concentra­
tions. For both species, the correlation between THg and MeHg was
greater than 97%. Positive and significant correlations were found be­
tween THg, MeHg and As concentrations for both species,.
Results of the correlation between condition factor K, THg, MeHg
and As concentrations in muscle tissue of fish species with genotoxic
variables (MN, nuclear buds, binucleated) are showed in Table 4. Posi­
tive correlations were found between cytogenetic variables and THg and
MeHg concentrations, finding significant values between MN, nuclear
buds and binucleated cells. Likewise, positive correlation was found for
As. According to the correlation coefficient between the K value and the
frequency of micronuclei, nuclear buds and binucleated cells there are
an inverse relationship between those variables, i.e., as the K value in­
creases, all frequencies decreases.
P. magdalenae
Factor K Total length (cm) Weight (g) Factor K
0.95 ± 0.15 25.2 ± 2.4 187.7 ± 50.8 1.15 ± 0.08
0.85 ± 0.16 23.9 ± 2.1 165.6 ± 33.3 1.22 ± 0.12
0.91 ± 0.11 28.4 ± 4.8 292.8 ± 42.7 1.42 ± 0.53
1.01 ± 0.14 25.7 ± 2.0 272.6 ± 19.9 1.65 ± 0.31
Á. Cruz-Esquivel et al. Ecotoxicology and Environmental Safety 253 (2023) 114670

Fig. 2. THg, MeHg and As concentrations (µg/kg) in fish dorsal tissue samples of (a) H. malabaricus and (b) P. magdalenae. Different letters indicate statistically
significant differences. Kruskal Wallis test with Dunn’s correction was used to identify the differences among groups.

5
Á. Cruz-Esquivel et al. Ecotoxicology and Environmental Safety 253 (2023) 114670

Table 2 Mercury has strong toxicological effects on the cell membrane as a result
Frequency of micronuclei and other nuclear alterations recorded in erythrocytes of its ability to cross membranes and impair cell metabolism systems
of P. magdalenae (PM) and H. malabaricus (HM). Data are represented as means of (AnvariFar et al., 2018). The Hg concentrations found in this research
eight samples ± SE. were significantly high, representing an ecological risk to the ecosystem.
Control Mojana 1 Mojana 2 Bajo Cauca On the other hand, As is widely present in the earth’s crust in at least
PM 10.5 29.7 25.0 26.6 moderate abundance, and has a wide variety of industrial uses,
± 3.6b ± 14.2a ± 6.25a ± 10.6a including pesticides. The toxicity is a result of oxidative stress caused by
Micronucleus HM 9.1 17.7 20.4 20.8 their inorganic forms, and not by the organic ones (Kumari et al., 2017).
± 3.9b ± 7.8a ± 6.3a ± 9.8a Among all species of As, the inorganic form (iAs), including arsenite
MW- 0.97 0.062 0.90 0.85
test
(As-III) and arsenate (As-V) are the most toxic forms found in Group 1/A
PM 0 ± 0b 3.6 ± 1.5a 4.4 ± 1.1a 7.2 ± 2.3a considered to be a human carcinogen (IARC, 1987).
Nuclear buds HM 1.4 4.7 ± 1.9a 7.3 ± 4.8a 8.8 ± 2.9a The positive correlations found for P. magdalenae evidence that the
± 0.5b sources of Hg and As are similar, e.g. residues from gold mining activ­
MW- 0.93 0.30 0.007 * 0.82
ities. Pinedo-Hernandez et al. (2015) reported high MeHg concentra­
test
PM 1.1 6.4 ± 2.7 a
5.5 ± 2.1 a
12.6 tions in the aquatic ecosystems in this area, possibly due to methylation
± 0.3b ± 4.3a of Hg from mining. The negative correlations found for H. malabaricus
Binucleated HM 0 ± 0b 4.3 ± 1.3a 6.3 ± 3.3a 6.4 ± 1.7a suggest that there is an important source of As other than mining, that
MW- 0.99 0.09 0.42 0.087 may come from an excessive use of agrochemicals that are also a source
test
of As (Cuenca et al., 2019; Naccarato et al., 2020).
MW-test = Mann Whitney values between H. malabaricus (HM) and P. magdalenae is an iliophage habit species located in a very low
P. magdalenae (PM) for each parameter in each site. position in the aquatic food chain, as opposed to H. malabaricus, a
Means with the same superscript letter are not significantly different from each carnivorous fish that presented higher Hg and As levels in its tissues.
other (P < 0.05). This bioaccumulation may possibly be influenced by many factors, such
as habitat, ecological needs, feeding habits and the aquatic environment.
Both elements have negative toxic effects on fish (Smylie et al., 2016;
Table 3
Spearman correlation P. magdalenae and H. malabaricus *Correlation signifi­ Kumari et al., 2017), such as the immune system, inducing pathological
cance at p < 0.05, * *Correlation significance at p < 0.01. changes among others associated with contamination by toxic elements
(Javed and Usmani, 2015).
Length Weight Factor K Hg MeHg
In our study, based on the analysis of MN and other nuclear abnor­
P. magdalenae: malities in peripheral blood erythrocyte cells of fish, a clastogenic and
Weight 0.801 * *
cytotoxic damage in the ictic species was evidenced. The concentrations
Factor K -0.593 * * -0.133
Hg 0.534 * * 0.274 * * -0.726 * * of Hg and MeHg, found in muscle tissue of fish species, were positively
MeHg 0.533 * * 0.279 * * -0.718 * * 0.968 * * associated with a genotoxic effect on red blood cells. Similar studies with
As 0.082 -0.256 * * -0.487 * * 0.447 * * 0.468 * * Channa punctatus (Yadav and Trivedi, 2009) showed a significant in­
H. malabaricus:
crease in micronuclei, with an order of increasing frequency of toxicity
Weight 0.923 * *
Factor K -0.820 * * -0.581 * *
and micronuclei of As > Hg; and Porto et al. (2005) evaluated the
Hg 0.542 * * 0.365 * * -0.620 * * genotoxicity in three fish species in the Madeira River in the Amazonas
MeHg 0.530 * * 0.357 * * -0.605 * * 0.994 * * basin impacted by gold mining activities and showed a high frequency of
As -0.094 -0.300 * 0.173 0.264 * * 0.25 * * micronuclei, especially in carnivorous fish (e.g. H. malabaricus),
In general, fish populations in the three studied impacted areas
showed significantly higher values for MN and ENA compared to the
Table 4 control group. The highest frequencies of MN were recorded in the
Spearman correlation (r-value) of Hg, MeHg and As concentrations (μg/kg), Mojana 1 site, probably due to a chronic effect of exposure to Hg con­
factor K and genotoxic parameters in fish species. *Correlation significance at centrations that drain with the floods from the Nechi and Cauca rivers.
p < 0.05, * *Correlation significance at p < 0.01. These results are consistent with those found in tropical carnivorous fish
Factor K Hg MeHg As (Coimbra et al., 2018), suggesting that Hg can cause aneugenicity and
P. magdalenae: clastogenicity affecting mainly mitotic division.
Micronucleous -0.465 * * 0.498 * * 0.454 * * 0.230 * * Although the Mojana 1 and 2 sites showed lower THg and MeHg
Nuclear buds -0.497 * * 0.546 * * 0.518 * * 0.416 * * concentrations than the Bajo Cauca, these concentrations were signifi­
Binucleated -0.212 * * 0.366 * * 0.342 * * 0.223 * * cantly higher compared to those found in the control group and other
H. malabaricus:
Micronucleous -0.448 * * 0.628 * * 0.618 * * 0.295 * *
studies. Likewise, the high frequencies of MN and ENA suggest that there
Nuclear buds -0.386 * * 0.636 * * 0.646 * * 0.235 * is an association with the presence of THg and MeHg in the tissues of
Binucleated -0.314 * * 0.570 * * 0.558 * * 0.410 * * both species. On the other hand, the Mojana 1 site showed high values of
As in fish tissues compared to the other sites. These high As levels may be
related to the nuclear alterations found in fish in this area, since the toxic
4. Discussion
effects of inorganic As can cause denaturation of cellular enzymes,
alteration of genetic regulation and inhibition of mitotic division
Historically, the regions of Bajo Cauca and Mojana have shown
delaying the correct assembly of the mitotic spindle. Singh et al. (2019)
contamination processes caused by mining activities that have deterio­
demonstrated the significant genotoxic effect of As bioaccumulation in
rated the quality of fishing resources (Marrugo-Negrete et al., 2008,
C. punctatus cells, finding a high frequency of micronuclei and nuclear
2018; Pinedo-Hernández et al., 2015). Previous research in the Mojana
alterations.
basin has documented high Hg concentrations in water ecosystems
On the other hand, positive and significant correlations were found
(Marrugo-Negrete et al., 2010, 2019; Pinedo-Hernández et al., 2015),
between Hg and MeHg concentrations with the individual length and
since it has been widely used in artisanal gold mining in the amalgam­
weight (Table 3). However, significant negative correlations were
ation process (Calao-Ramos et al., 2021). This metal is bioaccumulated
recorded between the condition factor K and the Hg, MeHg and As (no
and biomagnified by aquatic organisms throughout the food chain.
significant) concentrations in the dorsal tissue of both species. The K is

6
Á. Cruz-Esquivel et al.
generally used as an initial screening biomarker and can be affected by
several non-pollutant factors (Linde-Arias et al., 2008). Fish with low K
may indicate low fat deposit due to reduced food availability and/or
increased physiological demands for energetic resources (Jonge et al.,
2015). Decreasing in the K is also associated to lower foraging activity in
cold periods and to the end of the reproduction period (Gomiero and
Braga, 2005). The condition factor K, provides information on the
growth rate and feeding behavior of the fish studied. A value greater
than 1 cm/g indicates the welfare and robustness of the fish (Vasan­
tharajan et al., 2014). The individuals’ H. malabaricus were found with a
lower index than 1, probably because their carnivorous dietary habit has
caused them to bioaccumulate high metal loads in muscle tissue while
the P. magdalenae species were in a better condition, with values above 1
(Table 1). Similar results for Abramis brama L species fish were reported
by Farkas et al. (2003), that present high Hg concentrations in muscle
tissue and negative correlations with condition factor K. These data
provide fundamental information on growth strategies and nutritional
status of ictic species. In addition, these parameters are widely used to
compare the condition of populations that inhabit aquatic systems with
a high anthropogenic impact. Overall, the results in our study indicated
that the evaluated fishes attain a better condition (high K) when the
THg, MeHg and As concentrations are low. Moreover, the increase in
THg, MeHg and As, and decrease of K indicates rearrangement of energy
due to a detoxification mechanism. This also suggest that the depleting
reserves were on an upward growth due to the bioaccumulation of MeHg
(Erasmus et al., 2019). Even though K can be used to elucidate the effect
of contaminants on fish health, this should however be interpreted
cautiously because various factors such as sex, population, prey quality
and quantity consumed, season, or differences in environmental condi­
tions play an important role on the condition factor (Froese, 2006;
Martyniuk et al., 2020; Gutiérrez-Mosquera et al., 2021).
Studies related with condition factor and genotoxic variables are
very limited, however, literature provide some research that leads to a
close relationship between the frequencies of MN and nuclear abnor­
malities, with the condition factor K. Table 4 show a negative rela­
tionship between genotoxic variables with the K factor. Our results agree
with previous studies that revealed a decrease in K associated with the
high frequency of MN on the species Oreochromis niloticus and Mugil
cephalus (Omar et al., 2012), and also on Channa striatus and Hetero­
pneustes fossilis (Fatima et al., 2014). Likewise, Dalzochio et al. (2017)
reported a low condition factor K (0.98 ± 0.3) with high frequencies of
MN (0.31 ± 0.2) and ENA (3.48 ± 3.6) in fish from the Ilha River
(Brazil) compared to the control site. The species H. malabaricus regis­
tered high frequencies of MN and the lowest values of condition factor
(< 1), these results can be explained due to the feeding habit of this
species and a low capacity for migration. Finally, in a similar study using
H. malabaricus on the coast of the Imboassica swamp (Brazil) affected by
anthropic activities, fish species presented low values of condition factor
and high percentages of micronuclei in erythrocytes compared to con­
trol site (Coimbra et al., 2018).
5. Conclusions
The results reported in this study indicated that the Mojana and Bajo
Cauca areas have been impacted by anthropogenic activities due to gold
mining. Mercury, methylmercury and arsenic concentrations found in
fish, and their potential genotoxic effects on native aquatic species of
different dietary habits, can lead to a loss of aquatic diversity. The
species H. malabaricus and P. magdaneae registered a high genotoxic
impact on erythrocyte cells, and these abnormalities presented a nega­
tive correlation with the concentrations of Hg and MeHg, especially in
the Bajo Cauca region, which registered the highest concentrations of Hg
species. On the other hand, H. malabaricus registered low indices of
condition factor K. In addition, according to the high values of toxic
compounds found in edible fish its consumption may require attention
by local authorities due to adverse effects on human health. In this sense,
7
Ecotoxicology and Environmental Safety 253 (2023) 114670
fish consumption patterns and public knowledge about advisories and
warnings should be examined, as well as awareness on the permisibles
levels of Hg and As in fish. Environmental and genotoxic monitoring
programs should be established not only for fish, but also for water
samples, sediments, and other wildlife species that inhabit the region.
CRediT authorship contribution statement
Ángel Cruz-Esquivel: Investigation, Formal analysis, Methodology,
Writing − original draft. Sergi Díez: Conceptualization, Supervision,
Formal analysis, Writing − review & editing, José Luis Marrugo-
Negrete: Funding, Conceptualization, Supervision.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Data availability
Data will be made available on request.
Acknowledgments
The authors received financial support from the University of
́ Colombia through projects FCB-01–17 and FCB-01-
Có rdoba, Monteria,
19 This work was partly funded by the CSIC through the project
iCOOP2021- COOPA20490. The authors would also like to acknowledge
the CYTED (Programa Iberoamericano de Ciencia y Tecnologiá para el
Desarrollo), for financing the MercuRed Network (420RT0007).
References
Abdel-Khalek, A.A., 2015. Antioxidant responses and nuclear deformations in freshwater
fish, Oreochromis niloticus, facing degraded environmental conditions. Bull.
Environ. Contam. Toxicol. v. 94, 701–708.
Agudelo-Echavarría, D.M., Olid, C., Molina-Pérez, F., Vallejo-Toro, P.P., Garcia-
Orellana, J., 2020. Historical reconstruction of Small-scale gold mining activities in
tropical wetland sediments in Bajo Cauca-Antioquia, Colombia. Chemosphere 254,
126733.
Ali, H., Khan, E., 2019. Trophic transfer, bioaccumulation, and biomagnification of non-
essential hazardous heavy metals and metalloids in food chains/webs—Concepts and
implications for wildlife and human health. Ecol. Risk Assess. 25 (6), 1353–1376.
Anbumani, S., Mohankumar, M.N., 2019. The piscine erythrocyte micronucleus cytome
assay. Micron Assay. Toxicol. 228–242.
AnvariFar, H., Amirkolaie, A.K., Jalali, A.M., Miandare, H.K., Sayed, A.E., Üçüncü, S.,
Romano, N., 2018. Environmental pollution and toxic substances: cellular apoptosis
as a key parameter in a sensible model like fish. Aquat. Toxicol. 204, 144–159.
APHA, 1998. Standard Methods for the Examination of Water and Wastewater, 20th
ed..,, 1998. American Public Health Association,, Washington, DC.
Barone, G., Dambrosio, A., Storelli, A., Garofalo, R., Busco, V., Storelli, M., 2018.
Estimated dietary intake of trace metals from swordfish consumption: a human
health problem. Toxics 6 (2), 22.
Calao-Ramos, C., Bravo, A.G., Paternina-Uribe, R., Marrugo-Negrete, J., Díez, S., 2021.
Occupational human exposure to mercury in artisanal small-scale gold mining
communities of Colombia. Environ. Int. 146, 106216.
Ç avas, T., Ergene-Go¨zu¨kara, S., 2005. Induction of micronuclei and nuclear
abnormalities in Oreochromis nilo- ticus following exposure to petroleum refinery
and chromium processing plant effluents. Aquat. Toxicol. 74, 264–271.
Chen, H., Wang, P., Du, Z., Wang, G., Gao, S., 2018. Oxidative stress, cell cycle arrest,
DNA damage and apoptosis in adult zebrafish (Danio rerio) induced by tris (1, 3-
dichloro-2-propyl) phosphate. Aquat. Toxicol. 194 37–45.
Coimbra, R.S.C., Mascarenhas, M.S., Saraiva, V.B., Santos, C.R., Lopes, R.M., Hauser-
Davis, R.A., Carvalho, C.E.V., 2018. Metal loads and biomarker suite responses in a
tropical carnivorous fish indicative of anthropogenic impacts in a Southeastern
Brazilian lagoon. Environ. Model. Assess. 190 (9), 564.
Córdoba-Tovar, L., Marrugo-Negrete, J., Barón, P.R., Díez, S., 2022. Drivers of
biomagnification of Hg, As and Se in aquatic food webs: A review. Environ. Res. 204,
112226.
Cordy, P., Veiga, M.M., Salih, I., Al-Saadi, S., Console, S., Garcia, O., Roeser, M., 2011.
Mercury contamination from artisanal gold mining in Antioquia, Colombia: The
world’s highest per capita mercury pollution. Sci. Total Environ. 410, 154–160.
Cuenca, J.B., Tirado, N., Barral, J., Ali, I., Levi, M., Stenius, U., Dreij, K., 2019. Increased
levels of genotoxic damage in a Bolivian agricultural population exposed to mixtures
of pesticides. Sci. Total Environ. 695, 133942.
Á. Cruz-Esquivel et al.
D’Agostini, F., La Maestra, S., 2021. Micronuclei in fish erythrocytes as genotoxic
biomarkers of water pollution: an overview. Rev. Environ. Contam. Toxicol. v. 258,
195–240. https://doi.org/10.1007/398_2021_76.
Dalzochio, T., Simões, L.A.R., de Souza, M.S., Rodrigues, G.Z.P., Petry, I.E.,
Andriguetti, N.B., da Silva, L.B., 2017. Water quality parameters, biomarkers and
metal bioaccumulation in native fish captured in the Ilha River, southern Brazil.
Chemosphere 189, 609–618.
de Souza Neto, H.F., da Silveira Pereira, W.V., Dias, Y.N., de Souza, E.S., Teixeira, R.A.,
de Lima, M.W., Fernandes, A.R., 2020. Environmental and human health risks of
arsenic in gold mining areas in the eastern Amazon. Environ. Poll. 265, 114969.
Díez, S., Esbrí, J.M., Tobias, A., Higueras, P., Martínez-Coronado, A., 2011. Determinants
of exposure to mercury in hair from inhabitants of the largest mercury mine in the
world. Chemosphere 84, 571–577.
Erasmus, V.N., Iitembu, J.A., Hamutenya, S., Gamatham, J., 2019. Evidences of posible
influences of methylmercury concentrations on condition factor and maturation of
Lophius vomerinus (Cape monkfish). Mar. Pollut. Bull. 146, 33–38.
Farkas, A., Salánki, J., Specziár, A., 2003. Age-and size-specific patterns of heavy metals
in the organs of freshwater fish Abramis brama L. populating a low-contaminated
site. Water Res 37 (5), 959–964.
Fatima, M., Usmani, N., Hossain, M.M., Siddiqui, M.F., Zafeer, M.F., Firdaus, F.,
Ahmad, S., 2014. Assessment of genotoxic induction and deterioration of fish quality
in commercial species due to heavy-metal exposure in an urban reservoir. Arch.
Environ. Contam. tox67 2, 203–213.
Fenech, M., 2000. The in vitro micronucleus technique. Mutat. Res. /Fundam. Mol.
Mech. Mutagen. 455 (1–2), 81–95.
Froese, R., 2006. Cube law, condition factor and weight–length relationships: history,
meta-analysis and recommendations. J. Appl. Ichthyol. 22, 241–253.
García-Medina, S., Galar-Martínez, M., Gómez-Oliván, L.M., Ruiz-Lara, K., Islas-
Flores, H., Gasca-Pérez, E., 2017. Relationship between genotoxicity and oxidative
stress induced by mercury on common carp (Cyprinus carpio) tissues. Aquat.
Toxicol. 192, 207–215.
Gomes, J.M.M., Ribeiro, H.J., Procópio, M.S., Alvarenga, B.M., Castro, A.C.S., Dutra, W.
O., 2015. What the Erythrocytic Nuclear Alteration Frequencies Could Tell Us about
Genotoxicity and MacrophageIron Storage? PLoS ONE 10 (11), e0143029. https://
doi.org/10.1371/journal.pone.0143029.
Gomiero, L.M., Braga, F.M.S., 2005. The condition factor of fishes from two river basins
in São Paulo state. Southeast Braz. Acta Sci. Biol. Sci. 27 (1), 73–78.
Gutiérrez-Mosquera, H., Marrugo-Negrete, J., Díez, S., Morales, G., Montoya, L.,
Jonathan, M.P., 2021. Mercury distribution in different environmental matrices in
aquatic systems of abandoned gold mines, Western Colombia: Focus on human
health. J. Hazardous Materials 404, 124080.
Hina, N., Riaz, R., Ali, U., Rafique, U., Malik, R.N., 2021. A quantitative assessment and
biomagnification of mercury and its associated health risks from fish consumption in
freshwater lakes of Azad Kashmir, Pakistan. Biol. Trace Elem. Res. v. 199,
3510–3526. https://doi.org/10.1007/s12011-020-02479-z.
IARC, 1987. Monographs on the evaluation of the carcinogenic risk of chemicals to
humans. Supplement F. Overall Evaluation of Carcinogenicity. International Agency
for Research on Cancer. World Health Organization,, Lyon, France.
Jaramillo-Villa, U., Jiménez-Segura, L.F., 2008. Algunos aspectos biológicos de la
población de Prochilodus magdalenae en las ciénagas de Tumaradó (Río Atrato),
Colombia. Act. Biol. 30 (88), 55–66.
Jasim, M.A., Sofian-Azirun, M., Yusoff, I., Rahman, M.M., 2016. Bioaccumulation and
histopathological changes induced by toxicity of mercury (HgCl2) to tilapia fish
Oreochromis niloticus. Sains Malays. 45 (1), 119–127.
Javed, M., Usmani, N., 2015. Impact of heavy metal toxicity on hematology and glycogen
status of fish: a review. Proc. Natl. Acad. Sci. India B 85 (4), 889–900.
Jonge, M., Belpaire, C., Van Thuyne, G., Breine, J., Bervoets, L., 2015. Temporal
distribution of accumulated metal mixtures in two feral fish species and the relation
with condition metrics and community structure. Environ. Poll. 197, 43–54.
Khan, S., Rauf, R., Muhammad, S., Qasim, M., Din, I., 2016. Arsenic and heavy metals
health risk assessment through drinking water consumption in the Peshawar District,
Pakistan. Hum. Ecol. Risk Assess. 22 (3), 581–596.
Kumari, B., Kumar, V., Sinha, A.K., Ahsan, J., Ghosh, A.K., Wang, H., DeBoeck, G., 2017.
Toxicology of arsenic in fish and aquatic systems. Environ. Chem. Lett. 15 (1),
43–64.
Lamas, S., Ferná ndez, J.A., Aboal, J.R., Carballeira, A., 2007. Testing the use of juvenile
Salmo trutta L. as biomonitors of heavy metal pollution in freshwater. Chemosphere
67, 221–228.
Linde-Arias, A.R., Inácio, A.F., Alburquerque, C., Freire, M.M., Moreira, J.C., 2008.
Biomarkers in an invasive fish species, Oreochromis niloticus, to assess the effects of
pollution in a highly degraded Brazilian River. Sci. Total Environ. 399, 186–192.
Maggi, C., Berducci, M.T., Bianchi, J., Giani, M., Campanella, L., 2009. Methylmercury
determination in marine sediment and organisms by Direct Mercury Analyser. Anal.
Chim. Acta 641 (1–2), 32–36.
Maldonado-Ocampo, J.A., Ortega-Lara, A., Usma, O., Galvis, V., Villa- Navarro, F.A.,
Vasquez, G., Prada-Pedreros, S., Ardila, R., 2005. Peces de los Andes de Colombia.
Instituto de Investigaciones y Recursos Biológicos Alexander von Humboldt. Bogotá
DC 346.
Marrugo-Negrete, J., Verbel, J.O., Ceballos, E.L., Benitez, L.N., 2008. Total mercury and
methylmercury concentrations in fish from the Mojana region of Colombia. Environ.
Geochem Health 30 (1), 21–30.
Marrugo-Negrete, J., Benítez, L.N., Olivero-Verbel, J., Lans, E., Gutierrez, F.V., 2010.
Spatial and seasonal mercury distribution in the Ayapel Marsh, Mojana region,
Colombia. ↱. Int. J. Environ. Res. Public Health 20 (6), 451–459.
Marrugo-Negrete, J., Pinedo-Hernández, J., Díez, S., 2015. Geochemistry of mercury in
tropical swamps impacted by gold mining. Chemosphere 134, 44–51.
8
Ecotoxicology and Environmental Safety 253 (2023) 114670
Marrugo-Negrete, J., Pinedo-Hernández, J., Díez, S., 2015. Geochemistry of mercury in
tropical swamps impacted by gold mining. Chemosphere 134, 44–51.
Marrugo-Negrete, J., Pinedo-Hernández, J., Díez, S., 2017. Assessment of heavy metal
pollution, spatial distribution and origin in agricultural soils along the Sinú River
Basin, Colombia. Environ. Res. 154 380–388.
Marrugo-Negrete, J., Enamorado-Montes, G., Durango-Hernández, J., Pinedo-
Hernández, J., Díez, S., 2017. Removal of mercury from gold mine effluents using
Limnocharis flava in constructed wetlands. Chemosphere 167, 188–192.
Marrugo-Negrete, J., Pinedo-Hernandez, J., Combatt, E.M., Bravo, A.G., Diez, S., 2019.
Flood-induced metal contamination in the topsoil of floodplain agricultural soils: A
case-study in Colombia. Land Degrad. Dev. 30, 2139–2149.
Marrugo-Negrete, J., Pinedo-Hernández, J., Paternina–Uribe, R., Quiroz-Aguas, L.,
Pacheco-Florez, S., 2018b. Spatial distribution and evaluation of environmental
pollution by mercury in the Mojana region. Colombia Rev. MVZ Córdoba 23 ((Supl):
7062-7075), 7062–7075.
Marrugo-Negrete, J.L., Ruiz-Guzmá n, J.A., Ruiz-Ferná ndez, A.C., 2018a.
Biomagnification of mercury in fish from two gold mining-impacted tropical marshes
in northern Colombia. Arch. Environ. Cont. Toxicol. 74 (1), 121–130.
Marrugo-Negrete, J., Vargas-Licona, S., Ruiz-Guzmán, J.A., Marrugo-Madrid, S.,
Bravo, A.G., Díez, S., 2020. Human health risk of methylmercury from fish
consumption at the largest floodplain in Colombia. Environ. Res 182, 109050.
Martyniuk, M.A.C., Couture, P., Tran, L., Beaupre, L., Power, M., 2020. Seasonal
variation of total mercury and condition indices of Arctic charr (Salvelinus alpinus)
in Northern Québec, Canada. Sci. Total Environ. 738, 139450.
Mechora, Š., Germ, M., Stibilj, V., 2014. Monitoring of selenium in macrophytes— the
case of Slovenia. Chemosphere 111, 464–470.
Mela, M., Randi, M.A., Ventura, D.F., Carvalho, C.E., Pelletier, E., Ribeiro, C.O., 2007.
Effects of dietary methylmercury on liver and kidney histology in the neotropical fish
Hoplias malabaricus. Ecotoxicol. Environ. Saf. 68 (3), 426–435.
Mojica, J., Castellanos, C., Usma, J., Álvarez, R., 2002. Libro rojo de peces dulceacuícolas
de Colombia. Serie Libros Rojos de Especies Amenazadas de Colombia. Universidad
Nacional de Colombia y Ministerio del Medio Ambiente. Bogotá 288.
Mojica, J., Valderrama, M., Barreto, C., Álvarez-León, R., 2012. Prochilodus magdalenae
Steindachner, 1878. In: Mojica, J.I., Usma, J.S., Álvarez-León, R., Lasso, C.A. (Eds.),
Libro rojo de peces dulceacuícolas de Colombia (2012) Instituto de Investigación de
Recursos Biológicos Alexander von Humboldt/Instituto de Ciencias Naturales de la
Universidad Nacional de Colombia/Colombia/Universidad de Manizales. Bogotá,
pp. 134–138.
Monteiro, D.A., Rantin, F.T., Kalinin, A.L., 2013. Dietary intake of inorganic mercury:
bioaccumulation and oxidative stress parameters in the neotropical fish Hoplias
malabaricus. Ecotoxicology 22, 446–456. https://doi.org/10.1007/s10646-012-
1038-5.
Naccarato, A., Tassone, A., Cavaliere, F., Elliani, R., Pirrone, N., Sprovieri, F., Giglio, A.,
2020. Agrochemical treatments as a source of heavy metals and rare earth elements
in agricultural soils and bioaccumulation in ground beetles. Sci. Total Environ. 749,
141438.
Obe, G., Pfeiffer, P., Savage, J.R.K., Johannes, C., Goedecke, W., Jeppesen, P.,
Natarajan, A.T., Martínez-López, W., Folle, G.A., Drets, M.E., 2002. Chromosomal
aberrations: formation, identification and distribution. Mutat. Res 504, 17–36.
Omar, W.A., Zaghloul, K.H., Abdel-Khalek, A.A., Abo-Hegab, S., 2012. Genotoxic effects
of metal pollution in two fish species, Oreochromis niloticus and Mugil cephalus,
from highly degraded aquatic habitats. Mutation Research/Genetic Toxicol and Environ
Mutagen, 746 (1), 7–14.
Palacios-Torres, Y., Caballero-Gallardo, K., Olivero-Verbel, J., 2018. Mercury pollution
by gold mining in a global biodiversity hotspot, the Choco biogeographic region,
Colombia. Chemosphere 193, 421–430.
Perera, P.C.T., Sundarabarathy, T.V., Sivananthawerl, T., Kodithuwakku, S.P.,
Edirisinghe, U., 2016. Arsenic and cadmium contamination in water, sediments and
fish is a consequence of paddy cultivation: evidence of river pollution in Sri Lanka.
Achiev in the Lif. Sci 10 (2), 144–160.
Pinedo-Hernández, J., Marrugo-Negrete, J., Díez, S., 2015. Speciation and bioavailability
of mercury in sediments impacted by gold mining in Colombia. Chemosphere 119,
1289–1295.
Porto, J.I., Araujo, C.S., Feldberg, E., 2005. Mutagenic effects of mercury pollution as
revealed by micronucleus test on three Amazonian fish species. Environ. Res 97 (3),
287–292.
Rashed, M.N., 2001. Monitoring of environmental heavy metals in sh from Nasser Lake.
Environ. Int 27, 27–33.
Romero-Suárez, D., Pérez-Flórez, L., Consuegra-Solórzano, A., Vidal-Durango, J.,
Buelvas-Soto, J., Marrugo-Negrete, J., 2022. Total mercury (T-Hg) in ichthyofauna
with the highest consumption in San Marcos - Sucre. Colomb. Rev. MVZ Córdoba
Vol.27 (3), e2488.
Seiyaboh, E.I., Harry, G.A., Izah, S.C., 2016. Length-Weight Relationship and Condition
Factor of Five Fish Species from River Brass. Niger. Delta Biotech. Res 2 (4), 187–192.
Singh, M., Khan, H., Verma, Y., Rana, S.V.S., 2019. Distinctive fingerprints of
genotoxicity induced by As, Cr, Cd, and Ni in a freshwater fish. Environ. Sci. Poll. Res
26 (19), 19445–19452.
Smylie, M.S., McDonough, C.J., Reed, L.A., Shervette, V.R., 2016. Mercury
bioaccumulation in an estuarine predator: Biotic factors, abiotic factors, and
assessments of fish health. Environ. Pollut. 214, 169–176.
Szkoda, J., Zmudzki, J., Grzebalska, A., 2006. Determination of arsenic in biological
material by hydride generation atomic absorption spectrometry method. Bull. Vet.
Inst. Pulawy 50 (2), 269–272.
Vasantharajan, M., Jawahar, P., Sundaramoorthy, B., Venkatasamy, M., 2014. Length-
weight relationship of Lethrinus lentjan (La Cepede, 1802) and Lethrinus nebulosus
Á. Cruz-Esquivel et al.
(Forsskal, 1775) exploited in Thoothukudi Coast, Tamil Nadu, India. Int J. Vet.
Anim. Sci. Res 43, 14–18.
WHO, 1990. Methylmercury in Environmental Health Criteria 101. World Health
Organization,, Geneva, p. 19 https://apps.who.int/iris/bitstream/handle/10665/
38082/9241571012_eng.pdf.
Ecotoxicology and Environmental Safety 253 (2023) 114670
Yadav, K.K., Trivedi, S.P., 2009. Sublethal exposure of heavy metals induces micronuclei
in fish, Channa punctata. Chemosphere 77, 1495–1500.
Yan, H., Li, Q., Yuan, Z., Jin, S., Jing, M., 2019. Research Progress of Mercury
Bioaccumulation in the Aquatic Food Chain, China: A Review. Bull. Environ.
Contam. Toxicol. 102 (5), 612–620.