Marine Pollution Bulletin 173 (2021) 112953

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Marine Pollution Bulletin

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Metal and metalloids concentration in Galapagos fish liver and

gonad tissues
Eduardo Franco-Fuentes a, Nicolas Moity b, Jorge Ramírez-González b, Solange Andrade-Vera b,
Dailos González-Weller c, Arturo Hardisson a, Soraya Paz a, Carmen Rubio a, Ángel J. Gutiérrez a, *
a
Department of Obstetrics and Gynecology, Pediatrics, Preventive Medicine and Public Health, Toxicology, Legal and Forensic Medicine and Parasitology, Spain
b
Charles Darwin Research Station, Charles Darwin Foundation, Santa Cruz, Galapagos, Ecuador
c
Canarian Public Health service, Central Laboratory, Santa Cruz de Tenerife, Spain

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

Keywords: The Galapagos Islands are one of the best-preserved archipelagos in the world. We sampled individuals of six
Galapagos Marine Reserve demersal and three pelagic species. The metal concentration of liver and gonad tissues were analyzed by
Metals Inductively Coupled Plasma Optical Spectrometry. Findings suggest none of the pelagic species showed metallic
Demersal fishes
concentrations that could pose a risk to their health or reproductive capacity.
Pelagic fishes
S. violacea and P. clemensi Zn levels were higher in liver than in gonads, indicating that these species could be
Fish liver
Fish gonads in their reproduction period. Zn risk toxicity for the study species is negligible. High concentrations of Cd were
found in the liver of C. princeps, C. affinis and P. albomaculatus which could cause morphological and physio­
logical alterations. We hypothesize that the high metal concentrations found in the demersal species could come
from the volcanism of the islands, since no major sources of anthropogenic metal contamination can be found in
the archipelago.

1. Introduction chain causing adverse effects and death in the biodiversity of aquatic
ecosystems. The bioaccumulation of metals depends on the species,
Metals and metalloids, whether of anthropogenic origin (e.g. in­ environmental conditions, and inhibitory processes. The rate of bio­
dustrial discharges, fossil fuels, etc.) or of natural origin (e.g. volcanic accumulation in aquatic organisms depends on the ability of the or­
activity, soil and rock erosion, etc.), are present in all aquatic ecosystems ganisms to metabolize these elements and their concentration in the
(Kojadinovic et al., 2007). Metallic contamination in aquatic environ­ water column (Mathias et al., 2014). Usually, carnivorous fish have
ments has been associated to toxic effects that metals and metalloids can more concentrations of heavy metals in their bodies because they are at
cause on biodiversity, such as population decline or loss of species the top of food chains. However, in some aquatic species, similar in size
(Araújo and Cedeño-Macias, 2016). In addition, metals can have a great to those in our study, higher metal concentrations were observed in
impact on aquatic ecosystems since they can bioaccumulate in organ­ omnivorous species, such as Cyprinus carpio, than in carnivorous ones
isms causing toxicity if metal levels are very high (Eroglu et al., 2015). such as Esox lucius (Ali and Khan, 2019).
Metals can accumulate in fish tissues and can cause toxicity by The Galapagos Islands are one of the best-preserved archipelagos in
influencing vital operations and reproduction of fish, weaken the im­ the world, with exceptional biodiversity (Governing Council of the
mune system, and induce morphological and pathological changes Galapagos Special Regime, 2015). The Galapagos Marine Reserve
(Authman et al., 2015). The degree of toxicity depends on several factors (GMR) was established in 1998 as a multiuse area, allowing small-scale
such as the exposure time, type of metal, species, tissues, and the artisanal fishing, tourism, and other activities. Tourism and fishing ac­
physicochemical parameters of the water (Eroglu et al., 2015). tivities depend on ecosystem services of the GMR. Tourism is the main
Furthermore, the liver is the target organ for the accumulation of metals economy activity and fishing is one of the main food production activity
due to its detoxification capacity (La Colla, 2016). in Galapagos (Governing Council of the Galapagos Special Regime,
Nevertheless, metallic elements can be bioaccumulated in the food 2015). It is important to investigate metal concentrations in key species

* Corresponding author.
E-mail address: [email protected] (Á.J. Gutiérrez).

https://doi.org/10.1016/j.marpolbul.2021.112953
Received 21 July 2021; Received in revised form 1 September 2021; Accepted 7 September 2021
Available online 16 September 2021
0025-326X/© 2021 The Authors. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/4.0/).
E. Franco-Fuentes et al. Marine Pollution Bulletin 173 (2021) 112953

that inhabit the GMR since it has been shown metals are adversely zones (~258 km2) where ~32,000 people live (DPNG, 2014; INEC,
affecting species or habitats in the Galapagos (Jimenez-Uzcategui et al., 2019).
2017) and may have health consequences on the local human popula­ The GMR is influenced by four currents, which exert different phe­
tion that consumes fish (Franco-Fuentes et al., 2021). nomena on the archipelago during the year. The Panama current marks
The only potential anthropic sources of metals could come from the the wet and warm season from December to June, while the Peru and
3% inhabited territory where human activities occur. However, the Cromwell currents mark the dry and cold season from May to November
Galapagos Islands are one of the most volcanically active areas in the (Palacios, 2004; Forryan et al., 2021) (Fig. 1). However, the South-
world, so the availability of metals from geological origin is potentially Equatorial current varies all year long. These currents allow for the
high (Kurz and Geist, 1999) and could bioaccumulate in Galapagos’ existence of a great biodiversity of species and marine ecosystems, and
fauna as it has been demonstrated for Galapagos waterbird species mark distinctive areas of influence differentiated in five bioregions: Far
(Jimenez-Uzcategui et al., 2017; Jiménez et al., 2020). North, Elizabeth, Western, Northern and Central-southeastern (Edgar
The aim of this research is to provide a baseline on the concentration et al., 2004).
of metals (heavy metals, microelements, macroelements and trace ele­
ments) in liver and gonads of consumable fish species in the Galapagos,
highlighting the possible effects on the health and reproductive capacity 2.2. Sampling
of these species.
We sampled hepatic and gonadal tissue between February and May
2. Material and methods 2019 from catch landings of fishers at Pelikan Bay dock in Puerto Ayora
(Santa Cruz Island), aiming for having an equal sex distribution for each
2.1. Study area species. The demersal species sampled were Ocean whitefish (Caulola­
tilus princeps), Galapagos sailfin grouper (Mycteroperca olfax), Mottled
The Galapagos Archipelago is located at approximately 1000 km scorpionfish (Pontinus clemensi), Whitspotted sand bass (Paralabrax
west of the coast of Ecuador. It comprises the Galapagos National Park albomaculatus), Misty grouper (Hyporthodus mystacinus) and Bighead
(~7985 km2 covering most of the terrestrial realm), the Galapagos tilefish (Caulolatilus affinis); while the pelagic species were Yellowfin
Marine Reserve (~138,000 km2 covering the waters 40 nm surrounding tuna (Thunnus albacares), Palm ruff (Seriolella violacea) and Wahoo
the islands) and the non-protected areas that covers the urban and rural (Acanthocybium solandri).
Following Takashima and Hibiya (1995), sex was determined from

Fig. 1. Study area with the marine bioregions according to Edgar et al. (2004) and Moity (2019), the limits of the main harbours and the location of the non-
protected areas, corresponding to urban and rural areas.

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the gonad colour, size, blood vessel density and internal and external 3. Results
texture. However, sex characterization was impossible for some in­
dividuals because they were underdeveloped. We did not identify males A detailed table with the average, standard deviation, median and
for M. olfax and females for P. albomaculatus. All individuals were interquartile range of the metal concentration obtained from demersal
weighted in fresh presentation (to the nearest gram) and measured and pelagic species can be found in Supplementary Material Table II.
(furcal length, FL, to the nearest cm) (Table 1). All the samples came The outcomes of all statistical tests of liver and gonads are available in
from the Central-southeastern bioregion of the GMR (Fig. 1). Supplementary Material Tables III and IV respectively.
Finally, samples were mobilized to the Charles Darwin Research
Station where they were standardized to 5 g (measured with decimal 3.1. Metal concentrations in pelagic species
precision scale Mettler Toledo SB16001), labelled in Ziploc bags, kept in
cold storage (≈− 18 ◦ C) and later mobilized to the Toxicology Area of the 3.1.1. Liver
University of La Laguna (Tenerife, Canary Islands, Spain) for processing Pelagic species (T. albacares, S. violacea and A. solandri) did not show
and subsequent analysis of Al, B, Ba Cd, Co, Ni, Pb, Sr, Ca, Cu, Cr, Fe, K, significant differences for Cr and Mg in liver; however, the concentra­
Li, Mn, Mo, Mg, Na, V and Zn. tions of Cd and Zn were statistically different between the three species
(Fig. 2).
2.3. Sample processing for metal concentration determination Moreover, the levels of Ca, Al, and Na did not show significant dif­
ferences between S. violacea and A. solandri, but both species showed
Liver and gonads samples were weighted (recorded with a Mettler differences with T. albacares. Furthermore, T. albacares and S. violacea
Toledo Classic Plus PB153-S/Fact analytical scales with milligram pre­ did not exhibit differences for Fe, Ni and Pb among themselves; how­
cision) before they were placed in porcelain crucibles and dried in the ever, both species had differences with A. solandri. The concentrations of
oven at 80 ◦ C for 24 h until constant weight. After that, samples were Cu and K were not different between T. albacares and A. solandri, but
placed in the muffle oven for 48 h at 450 ± 25 ◦ C until white or gray- these species showed differences with S. violacea.
white ashes were obtained (Lozano-Bilbao et al., 2018).
Ashes were weighted and filtered through a 1.5% nitric acid solution 3.1.2. Gonads
with double distilled water until a final solution of 25 mL was obtained. No significant differences were observed between the pelagic species
Once the samples were dissolved, they were introduced into sampling in the following elements: Al, Ca, Cr, Fe, Mg, Mn, Na and Zn (Fig. 2). Ni
jars for analysis and the metals were determined by Inductively Coupled concentration did not show significant differences between T. albacares
Optical Emission Plasma Spectrometry (ICP-OES) (Lozano-Bilbao et al., and S. violacea, but both differed with A. solandri.
2018).
Standard reference material, detection and quantification limits and 3.2. Metal concentration in demersal species
quality control for all elements analyzed by Inductively Coupled Plasma
Optical Spectrometry (ICP-OES) are available in Supplementary 3.2.1. Liver
Material. M. olfax and C. princeps showed significant differences in all elements
except K. Something similar happened between C. princeps and
2.4. Data analysis P. clemensi, which showed significant differences in all elements except
Al and K (Figs. 3 and 4).
First, we analyzed differences in metal concentrations for the nine On the one hand, significant differences were observed in M. olfax
species. Then we pooled the information and tested differences between with H. mystacinus in B, Ca, Co, Cr, Mg, Na, Sr, and Zn. On the other
demersal species (C. princeps, C. affinis, M. olfax, P. clemensi, hand, significant differences were also observed between C. princeps and
P. albomaculatus and H. mystacinus) and pelagic species (T. albacares, C. affinis in Cd, Mo, and Zn; and with H. mystacinus in all the elements
S. violacea and A. solandri). except Al, Co, and V.
Normality of the data was checked using graphical representation of
the data Kolmogorov-Smirnov and Shapiro-Wilk tests, and the homo­ 3.2.2. Gonads
geneity of the variances was checked using the Levene statistic (Zar, M. olfax and C. princeps showed significant differences in Al, Co, K, V
1999). Normality was systematically violated, so a Kruskal-Wallis (H. and Zn. However, M. olfax also had differences with P. clemensi in B, Mo,
test) non-parametric test was applied. In the case that H test showed Ni and Zn; with C. affinis in K and Mo; and with P. albomaculatus in Mo
significant differences, a Mann-Whitney U post hoc test was performed and Zn (Figs. 3 and 4). Furthermore, significant differences were
to check differences between pairs of data. A 95% confidence interval observed in C. princeps with C. affinis in Ca, Cu, Mn, and Mo.
was chosen for all the tests (Rubio et al., 2017a, 2017b). P. clemensi and C. affinis differed in Ca, Cr, Mn, and Ni. In addition,
C. affinis and H. mystacinus showed differences in Ba, Ca, Cd, Cu, Mn,
Mo, and Na. Finally, it is important to note that the only element in
which no significant differences were observed between any of the

Table 1
Length range (cm), total weight range (kg), sample size (n) and proportion of males, females and undetermined (UND) for liver and gonads samples per species.
Species Range of furcal length (cm) Range of total wet weight (w.w) (kg) Liver Gonads

n Male Female UND n Male Female UND

Thunnus albacares 52–131 2–35 35 12 12 11 28 11 11 6

Seriolella violacea 58–82 2–8 36 15 15 6 35 15 15 5
Acanthocybium solandri 127–169 13–27 10 3 2 5 6 3 2 1
Caulolatilus princeps 31–57 0.4–2 35 18 13 4 33 18 13 2
Caulolatilus affinis 29–39 0.3–0.6 7 3 3 1 6 3 3 0
Mycteroperca olfax 37–76 0.6–6 31 0 16 15 21 0 15 6
Pontinus clemensi 28–60 0.4–4 34 11 11 12 31 11 11 9
Hyporthodus mystacinus 47–150 3–26 13 1 4 7 6 1 4 1
Paralabrax albomaculatus 32–55 0.6–2 5 3 0 2 4 3 0 1

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E. Franco-Fuentes et al. Marine Pollution Bulletin 173 (2021) 112953

Fig. 2. Median with interquartile range for the metals studied (expressed in mg/kg wet weight) of pelagic species (T. albacares, S. violacea and A. solandri). Liver
tissues are shown in blue while gonad tissues are orange. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version
of this article.)

species studied was Pb. 3.3.2. Gonads

T. albacares and S. violacea showed differences with M. olfax,
3.3. Interspecific analysis (pelagic vs demersal) C. princeps, P. clemensi and C. affinis in their concentrations of Ca, Cr, Mg,
Mn, and Na. Furthermore, differences were also observed with the same
3.3.1. Liver species with S. violacea in Fe, with T. albacares in Al, and with A. solandri
For Cd and Co, S. violacea showed differences with all demersal in Ca. On the other hand, there are differences in the concentration of
species, while T. albacares differed from all but C. affinis and Mg between A. solandri and H. mystacinus; and, in the Na concentration
P. albomaculatus. In addition, A. solandri also differed with M. olfax and of A. solandri with M. olfax and with P. clemensi.
C. princeps in Cd (Figs. 2, 3 and 4). The Cd concentration in T. albacares showed differences with all the
T. albacares and A. solandri showed differences in Cu concentrations demersal species except for H. mystacinus, while S. violacea and
with all demersal species except for C. princeps and C. affinis. A. solandri differed with H. mystacinus for Cd and Co. In the case of Pb,
The K content of S. violacea differed from all demersal species except A. solandri did not show differences with any demersal species and
C. affinis; and T. albacares which also showed differences with M. olfax. T. albacares only differed with M. olfax and P. clemensi (Figs. 2, 3 and 4).
Furthermore, the three pelagic species showed differences in Mg with
C. princeps. The Zn concentration of A. solandri showed differences with
all demersal species, while S. violacea and T. albacares only differed with
P. clemensi and H. mystacinus (Figs. 2, 3 and 4).

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E. Franco-Fuentes et al. Marine Pollution Bulletin 173 (2021) 112953

Fig. 3. Median with interquartile range for the metals studied (expressed in mg/kg wet weight) of demersal species (C. princeps, C. affinis and M. olfax). Liver tissues
are shown in blue while gonad tissues are orange. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of
this article.)

4. Discussion However, it can be concluded that this implies a lower bioavailability

and/or a lower assimilation capacity of this element by this species.
4.1. Metal concentrations in pelagic species Our results showed lower concentrations of Cu (13.6 mg/kg w.w)
and Pb (0.1 mg/kg w.w) in T. albacares than those obtained by Sadeghi
4.1.1. Liver health implications et al. (2020) (31.8 and 2.13 mg/kg w.w, respectively) and those ob­
A. solandri showed the highest concentrations of Zn and Cu (118.9 tained by Dhaneesh et al. (2014) in the Lakshadweep Sea (Indian Ocean)
and 20.0 mg/kg w.w, respectively) of all the species studied. These high (20.5 and 0.2 mg/kg w.w, for Al and Cr, respectively). In addition, the
concentrations could be because the metallothioneins of this species, concentration of Cd, Pb and Ni in our study (11.0, 0.1 and 0.3 mg/kg w.
which are cysteine-rich metal-binding polypeptides that are important w, respectively) was higher than those found by Dhaneesh et al. (2014)
for Zn and Cu homeostasis, have a lower degree of activity, which will (0.4, 0.042 and 0.021 mg/kg w.w, respectively). It is important to
cause them to have a worse capacity to detoxification. High concen­ highlight the wide range of concentrations of the different elements that
trations of Cu in the liver could cause morphological and histological can be found in the Galapagos for T. albacares compared to what has
alterations (Authman et al., 2015). On the other hand, an excess of Zn been found in other parts of the world. Therefore, based on our results
could also be toxic and cause structural damage that affects the growth, Galapagos fish could have a lower bioabsorption capacity of Cu, Pb, Al,
development, and survival of the organism (Ali et al., 2014). However, Cr, Cd, Pb and Ni. However, it is difficult to establish the manifestation
the Zn concentrations of A. solandri and T. albacares (48.0 mg/kg w.w) of toxicity in individuals of this species since it can vary over time due to
obtained in this study were lower than those obtained by Sadeghi et al. environmental and feeding factors (Pandey and Madhuri, 2014). We
(2020) in the Oman Sea, in which concentrations of 239.5 mg of Zn/kg recommend continuing sampling and analyzing metals in liver for
w.w were observed in the liver of T. albacares. Due to the lack of in­ T. albacares to assess the levels and range of metal concentrations in the
formation on the dose-response relationship for Zn in Galapagos fish, we Galapagos.
cannot establish if this element affects the health of the species. Zn and Cd are known to present antagonism, since Cd competes for

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Fig. 4. Median with interquartile range for the metals studied (expressed in mg/kg wet weight) of demersal species (P. clemensi, H. mystacinus and P. albomaculatus).
Liver tissues are shown in blue while gonad tissues are orange. (For interpretation of the references to colour in this figure legend, the reader is referred to the web
version of this article.)

the active site of Zn, resulting in an inverse relationship between the individuals of this species showed a larger size (127–169 cm) than the
concentrations of these metals (Jakimska et al., 2011a, 2011b). This rest of the pelagic species, which would favor a greater bioaccumulation
phenomenon can be observed in S. violacea, which had the highest (Jakimska et al., 2011b).
concentration of Cd of all the species studied and had low concentrations Moreover, the levels of Ca, Al, and Na were lower in T. albacares than
of Zn. On the other hand, the high concentration of Cd could pose a risk in the rest of the species. This may be due, as mentioned before, to
to the species, since Cd is a non-essential element whose toxicity in differences in feeding patterns, environmental differences, or physio­
marine organisms can cause kidney damage, tumors, hypertension, liver logical differences between the species studied (Pandey and Madhuri,
dysfunction and decreased reproductive capacity (Ali et al., 2014). 2014). However, due to the limited information available for the species
Regarding pelagic species, A. solandri showed the highest levels of Fe A. solandri and S. violacea as well as for the genus Acanthocybium and
and Pb (117.3 and 0.6 mg/kg w.w, respectively). These elements Seriolella on the metals studied, a reliable comparison cannot be made.
accumulate preferentially in the liver since it is the target organ for their
accumulation (Jakimska et al., 2011b). The three pelagic species are top 4.1.2. Gonads health implications
predators, so the higher concentrations found in A. solandri could be due No significant differences were observed between the three pelagic
to differences in their feeding patterns. A. solandri has been documented species for Al, Ca, Cr, Fe, Mg, Mn, Na and Zn. However, it is important to
to feed on a wide range of species, collecting up to 70 different taxa from note that of the three species S. violacea presented the highest concen­
their stomachs (Zischke, 2012). However, studies performed in the GMR trations for most of these elements, particularly Zn (51.6 mg/kg w.w).
found that the main feeding source for A. solandri are pelagic fish The excess of this element could alter ovarian sterioidogenesis, i.e. the
(mainly jack mackerel - Trachurus sp.-and flying fish – Prognichthys biosynthesis of steroid hormones from cholesterol, through the modu­
tringa-) (Baque-Menoscal et al., 2012; Páez-Rosas et al., 2020). lation of heat shock proteins (HSPs), which play a vital role defending
On the other hand, the higher concentrations of the different ele­ the cells from toxicity (Moniruzzaman et al., 2020). However, the Zn
ments found in A. solandri could be due to the fact that the sampled concentration in the pelagic species of our study was lower than those

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obtained in Katsuwonus pelamis (Scombridae) by Pragnya et al., 2020 in seasonally due to factors such as the growth or reproductive period of
India (97.29 mg/kg w.w). Therefore, we cannot establish whether there the organisms (Ersoy and Çelik, 2010). On the other hand, the concen­
may be a risk related to Zn toxicity in pelagic species, so cytohistological trations of K, Na, Ca, and Mg shown by C. princeps and C. affinis in our
studies must be carried out to verify this. study are similar to those obtained in different demersal species by Ersoy
The liver is the target organ for Zn accumulation (Monikh et al., and Çelik (2010), so we assume that these elements are supporting the
2012). However, unlike the other pelagic species, S. violacea showed current state of health of this species.
higher concentrations of Zn in gonads than in liver. This could indicate C. affinis showed high concentrations of Al (51.9 mg/kg w.w), even
that this species is in its reproduction period, since during this process exceeding the values found in pelagic species. This result is even higher
the organisms need Zn for the sexual maturation of the gonads (Zn is than those obtained in the demersal species studied by Yilmaz et al.
used in the formation of sperm and eggs) (Moreno-Sierra et al., 2016). In (2010) in Iskenderun Bay (Turkey), who showed Al concentrations from
females, these metals are redistributed from the liver to the gonads 4.27 to 8.34 mg/kg w.w. Aluminum toxicity depends on the physico­
through vitellogenin, which is a complex glycophospholipoprotein chemical properties of the water (specially on its pH, being generally an
(Moreno-Sierra et al., 2016). In addition, vitellogenin can develop issue for freshwater fish), and it could cause respiratory problems or
functions as a carrier protein of metals as Ca, Fe, Cu and Mg, which are physiological alterations such as cardiovascular, haematologic or
incorporated to the oocytes (Mommsen and Walsh, 1988). endocrine disturbances (Authman et al., 2015).
The pelagic species in our study showed lower ranges of concentra­ M. olfax and P. clemensi presented similar concentrations in most
tions of Ni (0.0–0.2 mg/kg w.w), Cd (0.1–0.3 mg/kg w.w) and Pb (≃0.1 elements and lower than the rest of pelagic species, so we assume that
mg/kg w.w) compared to pelagic species studied by Anandkumar et al., the metallic concentrations found in these species do not pose any risk to
2018 in Borneo (1.3 to 4.3 mg/kg w.w for Ni, 0.6 to 2.7 mg/kg w.w for their health. However, it is important to highlight the concentrations
Cd, and 0.5 to 3.1 mg/kg w.w for Pb). Therefore, we consider that Ni, Cd shown by M. olfax for Cd and Pb (0.9 and 0.1 mg/kg w.w, respectively).
and Pb are less bioavailable for pelagic species of the Galapagos. These concentrations are much lower than those shown by the species
Mycteroperca fusca in the study carried out by Lozano et al. (2009) in the
4.2. Metal concentrations in demersal species Canary Islands (Spain), which showed values of 656.0 mg/kg w.w for
Cd, and 109.1 mg/kg w.w for Pb. These differences may be due to
4.2.1. Liver health implications environmental differences, including the degree of anthropization, or
Species of the genus Caulolatilus (C. princeps and C. affinis) generally feeding patterns (Pandey and Madhuri, 2014).
showed the highest metallic concentrations of all demersal species. The Finally, H. mystacinus generally presented the lowest metallic con­
biology of these species within the Galapagos is still poorly known, but centrations in most of the elements studied, so we consider that this
Elorduy-Garay and Caraveo-Patiño (1994) described the feeding species is the least susceptible to toxicity due to the metals studied.
behavior and the reproduction of C. affinis as similar to C. princeps. H. mystacinus is considered a top predator in the demersal habitat, but its
Therefore, the high concentration of metals could be due to the ab­ metallic concentrations do not coincide with the bioaccumulation pro­
sorption of metals from the sediments and/or due to differences in cess (Fishbase, 2021). This could be due to differences in their habitat or
feeding patterns with respect to the rest of demersal species (Naccari feeding habits, however, due to the little information available on the
et al., 2015). Fitch et al. (1971) established that the genus Caulolatilus ecology of this species in the Galapagos Archipelago, a reliable conclu­
feed on a great variety of invertebrates, preferring ostracods, which are sion cannot be made (Fishbase, 2021).
filter-feeders that easily absorb elements from the ocean environment.
Moreover, the Pb concentration in C. princeps and C. affinis was 4.2.2. Gonads health implications
higher than the rest of the demersal species, showing values of 0.3 and As in liver, the species of the genus Caulolatilus were those that
0.6 mg/kg w.w respectively. A high concentration of Pb can decrease the presented the highest metallic concentrations, especially C. affinis,
antioxidants of the cells, specifically Thiol-containing antioxidants, and which showed the highest concentrations for most elements. In addition,
cause a significant increase in the production of reactive oxygen species the high concentrations of Al found in C. affinis (51.9 mg/kg w.w) could
(ROS). All this can cause a situation of oxidative stress that can affect the cause high mortality due to early hatching and, in acidic media (which
health and reproduction of the organism (Authman et al., 2015). How­ could occur in the waters around the Galapagos in future), abnormal
ever, compared to demersal species studied by Yilmaz et al. (2010) in metabolisms during vitellogenesis, affecting oocyte maturation and
Iskenderun Bay, Turkey, who found liver Pb concentrations between spawning (Sandoval et al., 2017).
1.47 and 2.98 mg/kg w.w, Galapagos fish showed low levels of Pb. Moreover, C. princeps, as in liver, had the highest concentrations of
On the one hand, high concentrations of Cd were found in C. princeps macroelements. The order of these elements (K > Na > Ca > Mg) was the
(23.1 mg/kg w.w), P. albomaculatus (18.5 mg/kg w.w) and C. affinis same as that obtained by Yeltekin and Oguz (2018) in the gonadal tissue
(10.6 mg/kg w.w). These concentrations are higher than those obtained of Van fish (Alburnus tarichi). As with liver tissue, variations in the
in the demersal species studied by Ersoy and Çelik (2010) and by concentrations of these elements could occur due to biotic and/or
Türkmen et al. (2011), which showed Cd levels of 0.05-0.63 and 0.42- abiotic factors. However, we assume that the concentrations of these
0.88 mg/kg w.w, respectively. The volcanic activity of the archipelago elements found in our study are enough for a good state of health of the
could contribute to the natural inputs of Cd to Galapagos waters, and species and will not affect to the reproductive capacity of the organisms
could be affecting the health of these species (Raimundo et al., 2015). (Jezierska and Witeska, 2001).
This heavy metal can cause morphological and histological alterations, Cu and Mn are essential in cellular metabolism, but in high con­
affecting mortality, body size and different body morphometrics and centrations they could be extremely toxic (Gárriz et al., 2019). The range
deformities as vertebral curvatures (Authman et al., 2015). of Cu concentrations of the demersal species in our study (0.5-4.2 mg/kg
On the other hand, the species of the genus Caulolatilus showed the w.w) was similar to that obtained by Türkmen et al. (2011) (0.91 and
highest concentrations of macroelements, being Na > K > Ca > Mg, 2.17 mg/kg w.w). On the other hand, the Mn concentrations obtained in
while for the rest of the species the order was K > Na > Mg > Ca. Ersoy most species were between 0.1 and 0.4 mg/kg w.w, while C. affinis
and Çelik (2010) found that the usual order for these elements in the showed very high concentrations of this element (12.6 mg/kg w.w)
demersal species they studied was K > Na > Ca > Mg, however, varia­ compared to those obtained by Türkmen et al. (2011), whose concen­
tions in the accumulation of these elements may occur due to different trations did not exceed 2.81 mg/kg w.w.
factors such as length and body weight, and the physical-chemical pa­ Regarding Pb, it is important to note that it was the only element in
rameters of water (Jezierska and Witeska, 2001). In addition, the which no significant differences were observed between any of the
changes in the concentrations of these macroelements can vary species studied, observing ranges between 0.1 and 0.3 mg/kg w.w.

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E. Franco-Fuentes et al. Marine Pollution Bulletin 173 (2021) 112953

These concentrations are much lower than those observed by Türkmen, bioregion. Moreover, a cytohistological study should be carried out in
Türkmen, & Tepe in 2011 (0.51–0.7 mg/kg w.w) or Anandkumar et al., parallel to associate possible alteration at the cellular level in the liver
in 2018 (0.5–4.25 mg/kg w.w). Due to the lack of information on the and gonadal tissues to metal content. As previously stated, ideally,
dose-response relationship of Pb in the gonads of Galapagos fish, we future studies should aim at sampling a greater number of species and
cannot establish if this element affects the reproductive capacity of the locations, targeting both seasons, all bioregions and different life stages
species. Moreover, the low concentration of Pb found in the studied of the species. Mercury was intentionally left out from the analysis of
species could be established by a lower bioavailability of this element in this paper since a study focusing solely on mercury concentration is
the Galapagos waters. currently underway by the research team.
The species P. clemensi showed a similar pattern to S. violacea in the
accumulation of Zn since higher values were observed in gonads than in 5. Conclusions
liver. This could indicate that this species is in its reproduction period,
and they use Zn in the sexual maturation of the gonads (Moreno-Sierra This is the first study that thoroughly estimates the concentration of
et al., 2016). However, there is not much information on the life history metals (macroelements, microelements and trace elements) of fishing
of these species to confirm this hypothesis. species in Galapagos and performs an evaluation of the potential health
As in liver, H. mystacinus generally presented the lowest concentra­ implications of such metals on the species. Our data shows T. albacares
tions of all demersal species in many of the elements studied, so we do had a wide range of concentrations for the different elements. Zn levels
not consider that the reproductive capacity of this species may be of S. violacea and P. clemensi were higher in the liver (31.1 and 17.1 mg/
affected by metals toxicity. kg wet weight (w.w), respectively) than in the gonads (51.6 and 27.0
Demersal species showed concentrations of Cd ranging from 0.1 to mg/kg w.w, respectively), suggesting that these species could be in their
0.6 mg/kg w.w with, P. albomaculatus being the species that presented reproduction period.
the highest levels. However, in the study carried out by Türkmen et al. Species of the genus Caulolatilus (C. princeps and C. affinis) generally
(2011), the demersal species showed concentrations of Cd from 0.49 to showed the highest metallic concentrations in both tissues of all
1.25 mg/kg w.w. Thus, we consider that the Cd concentrations observed demersal species. Likewise, high concentrations of Cd were found in the
in our study will not cause any harm to the health and/or reproductive liver of the species C. princeps, C. affinis and P. albomaculatus (23.1, 10.6
capacity of the species. and 18.5 mg/kg w.w, respectively), which could cause morphological
and physiological alterations. On the one hand, the health of C. affinis
4.3. Uncertainty could be affected due to the high concentrations of Al found in its tissues
(51.6 mg/kg w.w). On the other hand, C. affinis also presented high
Our study is partially limited by not having a previous solid scientific levels of Mn in gonads (13.6 mg/kg w.w) that could affect their repro­
database about metals in liver and gonads of fishes in the Galapagos ductive capacity.
Marine Reserve. Even though our study is based on a relatively small An antagonism between the Zn and Cd concentrations was observed
sample size, other studies use a similar sampling size (Staudinger, 2011). in demersal and pelagic species, associated with possible differences in
Also, the samples analyzed in this study come from one bioregion only. the activity of their metallothioneins. In addition, the lower metal
Furthermore, for M. olfax and P. albomaculatus we only sampled females concentrations in H. mystacinus do not appear to agree with the bio­
and males, respectively. Future sampling should include males and fe­ accumulation process and may be due to differences in the absorption of
males, respectively. However, males of M. olfax are difficult to obtain metals from the ocean environment and/or differences in eating habits.
due to the monandric protogynous hermaphrodite nature of this species. Finally, authors hypothesize that the high metal concentrations found in
We assume that the metallic content found in fishes comes from the the demersal species could come from the volcanism of the islands, since
volcanic substrate of the archipelago, however, it is possible that the no major sources of anthropogenic metal contamination can be found in
currents affect the metallic concentrations in the food web. For instance, the archipelago.
it has been described that the Cromwell upwelling on the west of Isabela Supplementary data to this article can be found online at https://doi.
Island adds important quantities of iron to the water column (Palacios, org/10.1016/j.marpolbul.2021.112953.
2004). Metal content in tissues can vary seasonally, so our study is
biased since we only have samples from the warm season. However, CRediT authorship contribution statement
since all samples were collected in the Central-southeastern bioregion,
the influence of the Cromwell upwelling is expected to be minimal. In Nicolas Moity: Methodology, Investigation, Writing – original draft.
the future, samples should be collected in both the cold and dry season to Jorge Ramírez-González: Methodology, Investigation, Writing – orig­
test the importance of season variation in metal concentrations and inal draft. Solange Andrade-Vera: Methodology, Writing – review &
extend the sampling to the other bioregions of the GMR. As it has been editing. Dailos González-Weller: Validation, Resources. Arturo Har­
shown in other parts of the world (Farkas et al., 2003), different stages of disson: Methodology, Writing – review & editing. Soraya Paz: Inves­
the life cycle of the species imply changes, potentially having a direct tigation, Writing – review & editing. Carmen Rubio: Conceptualization,
impact on the metallic content. In order to address this, future studies Resources, Writing – review & editing. Ángel J. Gutiérrez: Conceptu­
should aim at sampling different life stages of the studied species. alization, Methodology, Software, Formal analysis, Investigation,
Finally, it is difficult to compare our results with studies from other parts Writing – review & editing, Supervision.
of the world due to the unique characteristics of the archipelago.
Therefore, long-term research is necessary to obtain reliable results. In Declaration of competing interest
this sense, this study can be considered as the baseline for metal bio­
accumulation studies in fish liver and gonads in the Galapagos. The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
4.4. Future research the work reported in this paper.

Determining the metal content in liver and gonads of fishes is the first Acknowledgements
step to increase knowledge about the health of populations, as well as
the potential damage to their reproductive capacity. In future studies, We would like to thank to the Charles Darwin Foundation and the
geographical information on the catches will be needed to check if there Galapagos National Park for their help in this study. We owe a special
are spatial differences in the metallic concentrations of the species per thanks to artisanal fishers Luis Bonilla, Jaime Ascencio, Jonathan Erazo

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E. Franco-Fuentes et al. Marine Pollution Bulletin 173 (2021) 112953

and Nelson Ibarra who kindly contributed with samples for this study. mendiculus, flightless cormorant Phalacrocorax harrisi, and waved albatross
Phoebastria irrorata. Mar. Ornithol. 45, 159–163.
Furthermore, we would like to thank to Mr. Bearez for letting us use his
Kojadinovic, J., Potier, M., Le Corre, M., Cosson, R.P., Bustamante, P., 2007.
photos in the graphical abstract. The research was done under the Gal­ Bioaccumulation of trace elements in pelagic fish from the Western Indian Ocean.
apagos National Park Directorate permit PC-41-20. Finally, we would Environ. Pollut. 146 (2), 548–566.
like to thank the Gordon and Betty Moore Foundation for their support Kurz, M.D., Geist, D., 1999. Dynamics of the Galapagos hotspot from helium isotope
geochemistry. Geochem. Cosmochim. 63, 4139–4156.
in funding this research. This publication is contribution number 2395 La Colla, N.S., 2016. Bioacumulación de metales en peces marinos y su distribución en
of the Charles Darwin Foundation for the Galapagos Islands. columna de agua, bajo diferentes gradientes en el estuario de Bahía Blanca.
Universidad Nacional del Sur, Argentina. Tesis Doctoral.
Lozano, G., Brito, A., Hardisson, A., Gutiérrez, Á., González-Weller, D., Lozano, I.J.,
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