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Iron and Manganese Retention of Juvenile Zebrafish (Danio rerio)
Exposed to Contaminated Dietary Zooplankton
(Daphnia pulex)—a Model Experiment
Petra Herman
1
&Milán Fehér
2
&Áron Molnár
2
&Sándor Harangi
1
&Zsófi Sajtos
1
&László Stündl
2
&István Fábián
1
&
Edina Baranyai
1
Received: 15 January 2020 /Accepted: 10 May 2020
#The Author(s) 2020
Abstract
In present study the effect of iron (Fe) and manganese (Mn) contamination was assessed by modeling a freshwater food web of
water, zooplankton (Daphnia pulex), and zebrafish (Danio rerio) under laboratory conditions. Metals were added to the rearing
media of D. pulex, and enriched zooplankton was fed to zebrafish in a feeding trial. The elemental analysis of rearing water,
zooplankton, and fish revealed significant difference in the treatments compared to the control. In D. pulex the Mn level increased
almost in parallel with the dose of supplementation, as well as the Fe level differed statistically. A negative influence of the
supplementation on the fish growth was observed: specific growth rate (SGR%) and weight gain (WG) decreased in Fe and Mn
containing treatments. The redundancy analysis (RDA) of concentration data showed strong correlation between the rearing
water and D. pulex, as well as the prey organism of Fe- and Mn-enriched D. pulex and the predator organism of D. rerio.The
bioconcentration factors (BCF) calculated for water to zooplankton further proved the relationship between the Fe and Mn
dosage applied in the treatments and measured in D. pulex. Trophic transfer factor (TTF) results also indicate that significant
retention of the metals occurred in D. rerio individuals, however, in a much lower extent than in the water to zooplankton stage.
Our study suggests that Fe and Mn significantly accumulate in the lower part of the trophic chain and retention is effective
through the digestive track of zebrafish, yet no biomagnification occurs.
Keywords Trophic transfer .Iron .Manganese .Elemental analysis .Daphnia pulex .Danio rerio
Introduction
Aquatic ecosystems are considered to be the most sensitive
environmental media. Emitted pollutants tend to enter first to
surface waters, and through the aquatic food chain almost all
living organisms can be affected. Metal pollution of surface
waters is an ever increasing environmental issue of the last
few decades. Pollutants are released into the aquatic ecosystems
via anthropogenic activities (industrial and agricultural produc-
tion, traffic, untreated wastewater effluent etc.); however, the
natural geological background can also result in a higher
concentration of elemental impurities in the water body [1–3].
Metals accessing to water are usually the lack of biodegradabil-
ity compared to some organic materials that can be metabolized
into less harmful substances [2]. Beside the direct adverse effect
of the inorganic contamination on the aquatic plants and organ-
isms resulting in abnormalities, metals can be accumulated in
the cells and organs of living individuals [4–6]. Since the tro-
phic levels setting up the aquatic ecosystem are strictly built on
each other, more advanced and more complex organisms con-
suming species from the lower aquatic taxonomy class can
create much higher concentrations in their tissue trough bioac-
cumulation than it was in the initial environmental media [7].
Bioconcentration thus affects nearly all the living organisms in
the aquatic ecosystem as well as the human food chain [4,5],
which keeps the subject highly significant [3].
There are several factors on which the tendency and level
of bioaccumulation depend: from the distribution of the con-
taminants through the physical and chemical circumstances
(such as salinity, redox conditions, total suspended solid con-
centration) to the biology of the affected living organisms
*Edina Baranyai
baranyai.edina@science.unideb.hu
1
Department of Inorganic and Analytical Chemistry, Atomic
Spectroscopy Partner Laboratory, University of Debrecen,
Debrecen H-4010, Hungary
2
Faculty of the Agricultural and Food Sciences and Environmental
Management, University of Debrecen, Debrecen H-4032, Hungary
https://doi.org/10.1007/s12011-020-02190-z
/ Published online: 23 May 2020
Biological Trace Element Research (2021) 199:732–743
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
(feeding strategy, age, assimilation efficiency, etc.) [8,9].
Zhou et al. in their review considering the assessment of metal
pollution of aquatic ecosystems concluded that in order to gain
a deeper understanding of the pollution routes in aquatic me-
dia, further research of bioavailability is necessary [10]. The
condition of water ecosystems can be monitored by the con-
tinuous analysis of physical and chemical variables; however,
significant additional information can be gained by modeling
water-related pollution and investigating the effects on differ-
ent trophic levels by indicator organisms. Biomonitoring
plays an important role in this approach offering bioaccumu-
lation level and toxicological effects to support the compre-
hensive assessment [11]. Toxicity tests are carried out to in-
vestigate the effects of various chemicals and contaminants on
living organisms, comparing the sensitivity of the different
species [12–14]. They are most commonly applied to examine
the effect of the polluted media on the vitality, agility, and
survival of the indicator organisms [15]. However, by
bioindicators, it is also possible to investigate the accumula-
tion of certain pollutants along the food chain, thus gain more
detailed information regarding water quality [16–18]. In order
to examine the complex effect of more than one contaminant
and to evaluate the health condition of the entire aquatic eco-
system, it is important to consider the enrichment along the
trophic chain and to examine the biomagnification in the food
webs as well. Relationships within the life community can be
modeled by multispecies toxicity tests involving more taxa,
which are built on each other in the trophic system [19–22].
Both in active and passive aquatic toxicity studies bacteria,
algae, zooplankton, and fish species are most commonly ex-
amined [23]. These organisms represent different trophic
levels in the food chain and—depending on their
sensitivity—can provide information about the extent of the
pollution insurface waters. At the level of zooplankton organ-
isms, Daphnia pulex is one of the most common bioindicator
species of freshwaters due to the many characteristics facili-
tating its application in biochemical and toxicological studies
as well as its relatively simple and cheap raising under labo-
ratory conditions [14,15,24]. Daphnia species are considered
to be excellent indicators of the surrounding water, and being
at the base of the food chain, it is serving as resource for
consumers on higher trophic levels, including fish. Zebrafish
(Danio rerio) is on the next level in the trophic system and
also a very commonly used indicator organism [25,26]. Due
to its small size, rapid growth, ease of access, and other favor-
able biological features, individuals are often used not only in
toxicity tests but also in biomedical trials. Fernández et al.
mentioned in their very recent article that a steady increase
can be observed in the past 15 years of publications containing
“zebrafish”and “toxic”in the title, reaching an equal to that of
the mouse [27].
It is important to take into account the possible routes of
exposition when investigating the toxic effect of a chemical.
Depending on the form of the metal compound, uptake paths
are through the permeable epidermis or via food ingestion.
Since the entire body surface of the aquatic organism gets in
connection with the contaminants the adverse effects on the
outer epidermis, the digestive tract and the respiratory system
may add [28].
According to Koivisto [29] the development of complex
test systems corresponding to real nature is needed to better
assess and monitor the aquatic environment, while Zhou et al.
suggested the establishment of a precaution system for metal
pollution including biomonitoring network. In the scope of
these approaches the deeper understanding of the metal reten-
tion through the food chain is important and can be supported
by model experiments under laboratory conditions [22,30,
31]. Field studies alone are not enough to distinguish between
dietary and water-based metal retention [32].
Several studies can be found in the scientific literature in-
vestigating the toxic metal retention along the aquatic food
web; however, most of them are field studies of freshwater
ecosystems [33–37] and even more of them describe results
for marine environment [38–40]. Considering the analyzed
element, less data are available about Fe and Mn compared
to toxic metals such as Pb, Cd, As, or Hg [32,41–44].
Accordingly, toxic metal concentrations are more strictly reg-
ulated. WHO standards for drinking water (2011) has no strict
guideline for either Fe or Mn [45] (previous guidelines were
discontinued in the latest editions), while USEPA (2012, EPA
822-S-12-001) states maximum levels of 50 μgL
−1
and
300 μgL
−1
, respectively. EC (1998/83/E) directive is the
same for Fe and 200 μgL
−1
for Mn. Regarding the surface
waters and the level of protection necessary for aquatic life,
water quality standards give no acute criteria for Fe but con-
tain chronic criteria of 1.000 μgL
−1
.ConsideringMn,neither
acute nor chronic criteria are stated.
Thus, the aim of the current study is to investigate the tro-
phic transfer and biomagnification of Fe and Mn along the
aquatic food chain by exposing the metals to juvenile zebrafish
via dietary Daphnia pulex. Both metals are important
micronutrients and essential for aquatic organisms until
reaching a certain concentration limit. The Fe level was report-
ed to show a continuously increasing pattern in Swedish and
Finnish surface waters over the last 10 years [46], as well as
Mn is considered to be an emerging contaminant in the aquatic
environment [47]. The Fe and Mn level of the Hungarian ox-
bows in the Upper-Tisza region is recently reported to be high
as a result of our previous assessment [48] since when we
started to conduct model experiments involving these ele-
ments. Based on the pollution index of sediment samples, the
studied oxbows were characterized by moderate levels of con-
tamination for Fe and Mn, since the mean geochemical con-
centration in the upper level (0–10 cm) of floodplain sediments
is exceeded for both elements (35.000 mg kg
−1
for Fe
2
O
3
and
1000 mg kg
−1
forMnO)[49][50]. In our first study the Fe and
Iron and Manganese Retention of Juvenile Zebrafish (Danio rerio) Exposed to Contaminated Dietary... 733
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Mn retention was investigated via the rearing water media [28],
while in this paper the results of accumulation tendency
through food ingestion is described. The elemental concentra-
tion of the indicator organisms was determined by microwave
plasma atomic emission spectrometry (MP-AES), which is a
new and cost-effective technique that can be adapted in the
routine analysis of metal pollution.
Materials and Methods
Animals
Animal handling and experimental procedures followed the
Directive 2010/63/EU of the European Parliament and of the
council on the protection of animals used for scientific pur-
poses (2010/63/EU, Official Journal of the European Union. L
276/33. 20.10.2010.)
Water conditions were the same for both the maintenance
and treatment and checked regularly. Parameters among the
aquaria were not significantly different (p> 0.05) and did not
affect the treatments. Dissolved oxygen (DO), temperature,
and pH were tested daily by HACH LANGE HQ30D. The
level of ammonia, nitrite, and nitrate were measured weekly
by spectrophotometry (HACH LANGE DR3900) according
to the related USEPA methods (NH
4
-N: USEPA NESSLER
METHOD, NO
2
-N: USEPA DIAZOTIZATION METHOD,
NO
3
-N: CADMIUM REDUCTION METHOD).
During the experimental period, the following average wa-
ter quality parameters were determined:
DO : 7:55 0:34 mg L−1
Water temperature : 22:65 0:91°C
pH : 8:61 0:09
NH3þ:0:18 0:07 mg L−1
NO2
−:0:07 0:11 mg L−1
NO3
−:12:66:1mgL
−1
Enrichment of Daphnia pulex with Fe and Mn
Daphnia pulex individuals were originally collected from a
pond then reared isolated and enriched under laboratory con-
ditions in a model system, which ensured optimal environ-
mental conditions for the culture of the zooplankton organ-
isms. The consisting 4-L volume plastic tanks were filled up
with continuously aerated tap water, which temperature was
kept at 22 °C and a 16–8h(light-dark)ofilluminationwas
provided. A day prior to dietary serving the zooplankton, 0.6 g
wet mass of Daphnia was measured into each of the fifteen
plastic containers and enriched for 24 h according to the fol-
lowing treatments:
1. Fe: 5.70 mg L
−1
+ Mn: 2.90 mg L
−1
2. Fe: 5.70 mg L
−1
+ Mn: 6.25 mg L
−1
3. Fe: 15.0 mg L
−1
+ Mn: 2.90 mg L
−1
4. Fe: 15.0 mg L
−1
+ Mn: 6.25 mg L
−1
control, no supplementation
Concentrations were adjusted considering our preliminary
study where the retention was investigated via rearing water
[28]. In this work ten times the previously applied sublethal
level of Fe and Mn was adjusted to investigate the potential
accumulation effect of a toxic concentration range via the
chosen aquatic food chain.
The solutions of solid FeCl
3
and MnCl
2
(analytical purity,
SPEKTRUM 3D) were used to adjust the aforementioned
concentrations in the model media. Control treatment
contained only tap water with the elemental content of the
following: Cu: 7.0 μgL
−1
, Fe: 5.0 μgL
−1
,K:2.70mgL
−1
,
Mg: 16.1 mg L
−1
, Mn: 2.0 μgL
−1
, Na: 31.6 mg L
−1
, Sr:
0.40 mg L
−1
, and Zn: 41 μgL
−1
, according to ICP-OES anal-
ysis. Each treatment was set in triplicate(n= 3), and the plastic
containers were arranged in a completely randomized design.
After the enrichment period, the harvested D. pulex organisms
were filtered by plankton net of 150-μm mesh size and rinsed
with ultrapure water (Millipore MilliQ) in three times to evade
contamination of the rearing media. Daphnia pulex was fed ad
libitum to the zebrafish juveniles.
Enrichment of Danio rerio with Fe- and Mn-
Contaminated Daphnia
Zebrafish juveniles of 60dph were purchased from a local fish
market, and a 48 h of acclimatization period was applied at
25 °C prior to the feeding trial. The five treatments in triplicate
were arranged in rectangular glass aquaria of 40 L in a
completely randomized design with 10 zebrafish juveniles
(5–5 male and female) in each. The initial individual wet body
weight was 0.260 ± 0.035 g, and size homogeneity was tested
by ANOVA where no significant difference (p> 0.05) oc-
curred among the treatments. Aquaria were filled up with
aerated tap water; thus, the oxygen concentration was main-
tained at 100% during the 14 days of feeding trial. A 16–8h
(light-dark) of illumination was provided. Each aquarium was
filtered individually, and the flowing as well as aeration of
water was provided by piped filters.
The accumulation process of D. pulex was replicated daily;
thus, zebrafish juveniles were fed freshly enriched zooplank-
ton the same time every morning during the trial without ad-
ditional supplementation. The amount of zooplankton intro-
duced into the tanks was adjusted to obtain complete
Herman et al.
734
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
consumption. A 50% of water exchange took place daily as
well as aquaria were checked for dead individuals.
After the 14 days of enrichment period D. rerio individuals
were collected by fish net and were rinsed withultrapure water
to reduce the positive error in the analytical results. The indi-
vidual wet bodyweight was measured, and samples were kept
frozen prior to the sample preparation. The sacrificed proce-
dure was by physical methods suggested in the AVMA
Guidelines on Euthanasia for fish reported by the American
Veterinary Medical Association [51].
Sample Preparation and Elemental Analysis
Samples were dried at 105 °C for 24 h ina drying cabinet until
constant weight, and the dry body weight of the samples was
measured on analytical balance (Precisa 360 ES). They were
digested on an electric hot plate with 6.0 ml 65% (m/m) nitric
acid (reagent grade, Merck) and 2.0 ml 30% (m/m) hydrogen-
peroxide (reagent grade, Merck) at 80 °C for 4 h. After diges-
tion, samples were diluted with 1% (v/v) nitric acid (reagent
grade, Merck and Milli-Q water) to a final volume of 12 ml in
volume-calibrated test tubes.
Water samples from aquaria were collected every second
day to check the concentration of Fe and Mn: centrifuge tubes
of10ml(PP)withscrewcapswereusedforsamplingand1ml
of cc. HNO
3
was added to preserve until elemental analysis.
Elemental concentration was determined by microwave plas-
ma atomic emission spectrometer (Agilent MP-AES 4200).
Auto sampler (Agilent SPS4), Meinhard® type nebulizer and
double-pass spray chamber were used as well as a five-point
calibration procedure was applied (ICP VI, Merc). Certified ref-
erence material was used (ERM-BB422, fish muscle) to verify
that the measured elemental concentrations are equal with the
elemental levels of the examined organisms. The recoveries
were within 10% of the certified values for the metals. The
wavelengths and measuring parameters were chosen based on
the suggestion of the instrument’s software (MP Expert).
Data Evaluation
Weight gain (WG) percentage was calculated from the mea-
sured initial and final wet weight (W
i
and W
f
,respectively)
data of D. rerio individuals:
WG %ðÞ¼Wf−Wi
ðÞ=Wi100
Specific growth rate (SGR) was applied to describe the
growing performance of D. rerio according to the following
formula:
SGR %=dayðÞ¼lnWf−lnWi
ðÞ=t100;
where W
f
is the final wet body weight and W
i
is the initial wet
body weight of the zebrafish individuals [52].
The bioconcentration factor (BCF) for D. pulex was calcu-
lated by dividing the Fe and Mn concentration measured in the
zooplankton (C
tissue
,mgkg
−1
dry weight) by the same con-
centration values of the rearing water media (C
water
,mgL
−1
)
[28]:
BCF ¼Ctissue=Cwater
Trophic transfer factor (TTF) was calculated as the ratio of
the tissue concentration of Fe and Mn measured in D. rerio
(C
predator
) and the tissue concentration of the two elements
measured in D. pulex (C
prey
) both given in mg kg
−1
dry weight
[53]:
TTF ¼Cpredator=Cprey
The statistical evaluation of experimental data was carried
out in SpSS/PC+ software package. ANOVA was applied to
study the WG, SGR, BCF, TTF, and elemental concentration
results of the applied treatments. The homogeneity of variance
was checked by Levene test, and significant differences were
investigated by Tukey multi-comparison test where difference
wasconsideredtobestatistically proven when p<0.05.
Redundancy analysis (RDA) was performed in Canoco for
Windows 4.5 to study the interaction between the studied
species and their environmental background: D. pulex/water
and D. rerio/D. pulex,respectively.
Results
Survival and Growth Performance of Danio rerio Fed
with Fe- and Mn-Contaminated Daphnia
The survival of zebrafish juveniles was 100% in all treat-
ments; thus, the applied concentrations of Fe and Mn did not
cause the mortality of fish individuals during the experimental
period. The average final wet weight of fish increased in all
treatments compared to the initial values (Fig. 1)indicating
that the metal supplementation did not result in growth abnor-
mality; however, reduced growth performance was observed.
The highest percentage values of SGR and WG were achieved
by the control group (Table 1), which suggests that feeding
Fe- and Mn-enriched D. pulex might had a negative influence
on fish growth.
Elemental Concentration of Fe- and Mn-Enriched
Daphnia pulex and Danio rerio
The Fe and Mn concentration of D. pulex calculated to dry
weight is indicated in Fig. 2a and b, respectively.
Both the Fe and Mn level of the zooplankton samples in all
the applied treatments increased significantly compared to the
control group (p< 0.001, F=4.319 and F= 12.236,
Iron and Manganese Retention of Juvenile Zebrafish (Danio rerio) Exposed to Contaminated Dietary... 735
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
respectively). Considering the Mn content, the treatments con-
taining the same Mn supplementation (2.90 mg L
−1
in num-
bers 1 and 3 as well as 6.25 mg L
−1
in numbers 2 and 4) did
not differ statistically from each other (p> 0.05) (Fig. 2b).
Similar results for Fe were gained (Fig. 2a): its concentration
in treatment numbers 1 and 2 (both containing 5.70 mg L
−1
Fe) is statistically comparable (p> 0.05) as well as in treat-
ment numbers 3 and 4 (both containing 15.0 mg L
−1
)
(p> 0.05). However, in contrast to Mn, the Fe level differed
less between the treatments supplemented with the lower
(5.70mgL
−1
) and the higher (15.0 mg L
−1
) dosage
(p< 0.05, respectively).
The average Fe and Mn concentration of zebrafish juve-
niles per treatments are indicated in Fig. 2c and d, respective-
ly. The Fe content of all groups increased compared to the
control (F=4.013,p< 0.05); however, the two different ap-
plied Fe concentrations did not affect the Fe level of the
zebrafish individuals—no statistical difference was found ei-
ther between treatments 1 and 2 (5.70 mg L
−1
)(p>0.05) or
between treatments 3 and 4 (15.0 mg L
−1
)(p>0.05).TheMn
concentration of the fish also increased significantly compared
to the non-supplemented control group (F= 16.132,
p<0.001).
The elemental composition of D. rerio individuals can be
seen in Table 2. No significant difference occurred in the
composition of the samples regarding the trace element pat-
tern either compared to the control (p> 0.05, respectively, and
Ca: F=1.668;Cu:F=2.286;Mg: F=1.271;Na:F=0.895;
Zn: F= 0.887) or between the treatments, except for Co
(p<0.01, F= 37.326).
Interaction Between the Fe and Mn Levels of Water,
Daphnia pulex,andDanio rerio
Redundancy analysis was applied to assess the interaction
between the level of Fe and Mn in the treatments and in the
studied aquatic species, which is a commonly applied statisti-
cal technique to explain and model different cause–effect re-
lationships [54]. The RDA biplot regarding the Fe and Mn
concentration of the rearing water and D. pulex is indicated
in Fig. 3. In the first component (RDA1) the correlation be-
tween Fe and Mn concentration of the rearing media and the
concentration of the same elements in D. pulex was 0.884,
while in the second component (RDA2) the correlation was
0.858. The cumulative percentage variance of elemental con-
centration of the zooplankton was 77.2 (RDA1) and 78.1
Fig. 1 The average initial and
final wet weight of zebrafish
individuals in the different
treatments (mean ± SE, n=3)
Control: fed by non-
supplemented Daphnia,1:fedby
Daphnia supplemented with
5.70 mg L
−1
Fe and 2.90 mg L
−1
Mn, 2: fed by Daphnia
supplemented with 5.70 mg L
−1
Fe and 6.25 mg L
−1
Mn, 3: fed by
Daphnia supplemented with
15.0 mg L
−1
Fe and 2.90 mg L
−1
Mn, 4: fed by Daphnia
supplemented with 15.0 mg L
−1
Feand6.25mgL
−1
Mn
Table 1 Specific growth rate and weight gain percentage ofDanio rerio
in different treatments (mean ± SE, n= 3). Control: fed by non-
supplemented Daphnia,1:fedbyDaphnia supplemented with
5.70 mg L
−1
Fe and 2.90 mg L
−1
Mn, 2: fed by Daphnia supplemented
with 5.70 mg L
−1
Feand6.25mgL
−1
Mn,3:fedbyDaphnia
supplemented with 15.0 mg L
−1
Fe and 2.90 mg L
−1
Mn, 4: fed by
Daphnia supplemented with 15.0 mg L
−1
Fe and 6.25 mg L
−1
Mn.
Letters in lowercase indicate significant differences (p<0.05)
Treatments SGR (%/day) ± SE WG (%) ± SE
Control 2.20 ± 0.63a 37.0 ± 11.7a
1 1.15 ± 0.24b 17.5 ± 3.93b
2 1.42 ± 0.47c 22.6 ± 7.96bc
3 1.68 ± 0.52c 27.3 ± 9.01c
4 1.15 ± 0.26b 17.6 ± 4.25b
Herman et al.
736
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(RDA2). The relation regarding the species–environment con-
nection revealed to be 98.9 (RDA1) and 100.0 (RDA2). The
biplot shows a relatively strong correlation between the Fe
levels of the Fe- and Mn-contaminated water as well as the
reared zooplankton individuals, and a total agreement be-
tween the Mn concentrations.
Fig. 2 The Fe (a) and Mn (b) concentration of supplemented Daphnia
pulex and zebrafish (cand d, respectively) (mean ± SE dry weight, n=3)
Control: non-supplemented, 1: supplemented with 5.70 mg L
−1
Fe and
2.90 mg L
−1
Mn, 2: supplemented with 5.70 mg L
−1
Feand6.25mgL
−1
Mn, 3: supplemented with 15.0 mg L
−1
Fe and 2.90 mg L
−1
Mn, 4:
supplemented with 15.0 mg L
−1
Fe and 6.25 mg L
−1
Mn. Letters above
columns indicate significant differences (p<0.05)
Table 2 Elemental concentration (mg kg
−1
, dry weight) of Danio rerio
in different treatments (mean ± SE, n= 3). Control: fed by non-
supplemented Daphnia,1:fedbyDaphnia supplemented with
5.70 mg L
−1
Fe and 2.90 mg L
−1
Mn, 2: fed by Daphnia supplemented
with 5.70 mg L
−1
Feand6.25mgL
−1
Mn, 3: fed by Daphnia
supplemented with 15.0 mg L
−1
Fe and 2.90 mg L
−1
Mn, 4: fed by
Daphnia supplemented with 15.0 mg L
−1
Fe and 6.25 mg L
−1
Mn.
Letters in lowercase indicate significant differences (p<0.05)
Treatment Ca (mg kg
−1
)Co(mgkg
−1
)Cu(mgkg
−1
) Mg (mg kg
−1
)Na(mgkg
−1
) Zn (mg kg
−1
)
Control
N=30
Mean ± SE 29.346 0.346a 7.52 1046 2438 228
1864 0.0555 0.608 44.5 85.1 15.1
1
N=30
Mean ± SE 30.968 0.607b 8.63 1081 2449 245
2854 0.118 1.16 84.7 277 24.3
2
N=30
Mean ± SE 33.789 0.924c 9.75 1168 2637 255
3754 0.101 1.32 69.3 129 14.7
3
N=30
Mean ± SE 32.963 1.06c 7.99 1094 2369 244
2073 0.0623 0.715 52.8 252 11.4
4
N=30
Mean ± SE 33.714 1.09c 7.97 1108 2462 255
2518 0.0842 0.716 65.8 187 19.4
Iron and Manganese Retention of Juvenile Zebrafish (Danio rerio) Exposed to Contaminated Dietary... 737
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Very similar results were obtained for the RDA of the Fe
and Mn content of D. pulex reared in contaminated water
media and D. rerio fed by the enriched zooplankton organism,
shown also in Fig. 3. The correlation between Fe and Mn
concentrations in the relation of D. pulex and D. rerio in
RDA1 was 0.855, while in RDA2 the correlation was 0.987.
The cumulative percentage variances of Fe and Mn level of
the zebrafish were 72.4 (RDA1) and 73.3 (RDA2). The
species–environment connection was found to be 98.8
(RDA1) and 100.0 (RDA2).
Bioconcentration and Trophic Transfer Factor of
Daphnia pulex and Danio rerio
Bioconcentration factors calculated from water to zooplank-
ton for Fe and Mn are summarized in Table 3. Statistically
significant difference occurred (p<0.05,Fe:F= 21.727; Mn:
F= 3.407) between the treatments containing 5.7 mg L
−1
Fe
(treatments 1 and 2) and the treatments containing
15.0 mg L
−1
(treatments 3 and 4). Groups supplemented with
the same dosage of Fe did not differ from each other signifi-
cantly (p> 0.05). Higher supplemented level resulted in lower
BCF values. The same is true for Mn: the higher dose of
enrichment provided lower BCF proving the relation between
the measured concentration of the two elements applied in the
treatments and in the zooplankton organisms. The BCF data
for Fe in the treatments 1 and 2 are nearly four times to that of
the Mn. The TTF values are summarized in Table 4for
D. rerio exposed to dietary Fe and Mn via D. pulex,showing
less difference yet still statistically proven (p<0.05,Fe=F=
2.597; Mn: F=5.092).
Discussion
The accumulation tendency of different pollutants carries es-
sential information regarding the water quality, the well-being
Fig. 3 The RDA biplots of the Fe and Mn level of rearing water and D. pulex as well as of D. pulex and D. rerio (solid arrow: elemental concentration of
water/D. pulex, dashed arrow: elemental concentration of D. pulex/D. rerio. Filled circles with numbers indicate experimental groups)
Table 4 Trophic transfer factors (mean ± SE) for Fe and Mn from
Daphnia pulex to Danio rerio. Control: fed by non-supplemented
Daphnia, 1: fed by Daphnia supplemented with 5.70 mg L
−1
Fe and
2.90 mg L
−1
Mn, 2: fed by Daphnia supplemented with 5.70 mg L
−1
Feand6.25mgL
−1
Mn,3:fedbyDaphnia supplemented with
15.0 mg L
−1
Fe and 2.90 mg L
−1
Mn, 4: fed by Daphnia supplemented
with 15.0 mg L
−1
Fe and 6.25 mg L
−1
Mn. Letters in lowercase indicate
significant differences (p<0.05)
Elements Treatments TTF × 10
−3
Fe 1 7.19 ± 0.410a
2 6.74 ± 0.627a
3 5.51 ± 0.616b
4 5.15 ± 0.720b
Mn 1 4.45 ± 0.208a
2 3.26 ± 0.555b
3 3.26 ± 0.410b
4 2.23 ± 0.351c
Table 3 Bioconcentration factors (mean ± SE) of Fe and Mn from water
to Daphnia pulex. Control: non-supplemented, 1: supplemented with
5.70 mg L
−1
Feand2.90mgL
−1
Mn, 2: supplemented with
5.70 mg L
−1
Feand6.25mgL
−1
Mn, 3: supplemented with
15.0mgL
−1
Feand2.90mgL
−1
Mn, 4: supplemented with
15.0 mg L
−1
Fe and 6.25 mg L
−1
Mn. Letters in lowercase indicate
significant differences (p<0.05)
Elements Treatments BCF
Fe 1 4591 ± 105a
2 4371 ± 169a
3 1902 ± 31.1b
4 1864 ± 33.0b
Mn 1 1044 ± 54.3a
2 925 ± 96.3b
3 1006 ± 35.9a
4 799 ± 17.3b
Herman et al.
738
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
of aquatic ecosystems, and is directly related to human health.
The appearance of iron and manganese in water ecosystems
can originate from industrial and agricultural emissions; how-
ever, the natural geological background can result in elevated
levels.
The growth performance of zebrafish juveniles was found
to be slightly affected by the environmentally toxic levels of
Fe and Mn since lower values of SGR and WG were detected
compared to the control group. This finding is in contrast with
our previous experiment where the sublethal level of Fe and
Mn retention was studied from water via the gills of Common
carp (Cyprinus carpio)[28]. In that study ten times lower
level of the two elements were applied in a very similar ex-
perimental setup; thus, we can conclude that the elevated dose
of Fe and Mn supplementation in present investigation
reached the value where it affects the growth performance.
The negative influence of Mn in fish is mainly due to the
increased oxidative stress it provokes [55–58]. Gabriel et al.
described the toxicity of Mn exposed to juvenile Colossoma
macropomum by evaluating oxidative stress parameters and
found that biomarkers changed significantly in the tissues of
the studied fish individuals proving specific toxicity of Mn to
the different organs [58]. In contrast, low level of Mn supple-
mentation in fish diet can have opposite effect by promoting
growth and normal skeleton development [20].
The Mn level in the zooplankton organisms increased in
parallel with the dose of the applied supplementation. The
same phenomenon was observed by Fehér et al. in a 24-h
enrichment trial of Artemia nauplii with MnCl
2
[52]. The high
bioconcentration potential of mainly herbivores Daphnia spe-
cies is published compared to omnivorous species, such as
rotifers, for Cd, As, and Pb [33]. However, the two different
applied Fe concentrations in treatments did not result in sig-
nificantly different uptake level of the zebrafish individuals.
This finding correlates with the results obtained by Harangi
et al., who conduct experiments to study the accumulation of
metals in juvenile carp exposed to sublethal levels of Fe and
Mn [28]. Either the smaller accumulation tendency of Fe can
be one of the possible explanations or the higher hydrolyza-
tion potential of Fe in the model media in which the zooplank-
ton organism was reared. However, compared to the control
group, the concentration of both the studied elements in-
creased. This finding proves that nevertheless the main uptake
path for dissolved Fe and Mn for fish is via the gills; retention
is also efficient through the digestive track. Since the level of
the other essential elements did not change upon the Fe and
Mn treatments, we can further conclude that the applied con-
centrationof Fe and Mn did not affect the metabolism process
of other metals.
The good correlation indicated in the redundancy analysis
proves the direct connection between the Fe and Mn concen-
tration of the zooplankton organisms as well as the zebrafish
individuals with the applied treatments, thus the ability of
these elements to concentrate in the aquatic food chain.
Fehér et al. used RDA to reveal the effect between the ele-
mental content of a saltwater zooplankton (Artemia naupli)
species and Barramundi (Lates calcarifer)larvae[52]. In their
experimental setup the zooplankton was reared in a Mn, Zn,
and Co supplemented water media and barramundi individ-
uals were fed by the enriched Artemia. Similar to present
study, they found correlation between the environmental and
species concentration data for Mn.
Bioconcentration of organic and inorganic pollutants in
aquatic organisms is a significant parameter to assess the ef-
fect of chemicals emitted to the environment. It can be
expressed numerically in experiments conducted under labo-
ratory conditions by the bioaccumulation factor, which can be
used for regulatory purposes if certain limitations are consid-
ered [59]. The BCF of present study proved the relation be-
tween the level of Fe and Mn in the treatments and in the
zooplankton organisms. Manganese is a commonly occurring
element in the aquatic environment, in both underground and
surface waters. Although it is an essential element, even a
short-term waterborne exposure to higher concentration can
indicate peroxidative damage in fish tissues [60]. Marins et al.
in their fresh study from 2019 found that long-term exposure
to Fe and Mn in a concentration commonly found in ground-
water may cause damage to chromosome levels and changes
in locomotor and exploratory behaviors of adult zebrafish
[61]. It was further concluded by Tu H. et al. that develop-
mental exposure to Mn along with Cd and Pb statistically
reduced the velocity and distance of the larval swim of
zebrafish [62]. Altenhofen et al. studied the effect of MnCl
2
exposure on cognition and exploratory behavior in adult and
larval zebrafish. Both adults and larvae reacted with decreased
distance traveled and absolute body turn angle as well as in-
creased apoptotic markers were found in their nervous system.
The research group concluded that the prolonged Mn expo-
sure resulted in locomotor deficits that can damage the dopa-
minergic system [63].
The BCF results for Fe in the first two treatments of present
study are higher than the same data for Mn. In our previous
study the Fe and Mn bioconcentration was investigated from
water to fish (Common carp) in a model experiment and sim-
ilar tendency was found [28]. Voigt et al. also reported much
higher BCF for Fe compared to Mn in the tissue of Geophagus
brasiliensis originating from Alagados Reservoir, Ponta
Grossa [64]. Iron can affect both directly and indirectly the
living organisms in aquatic ecosystems and, according to the
suggestion to Vuori, the ecotoxicological aspects of Fe should
be further analyzed with organisms of different levels [65].
Liu et al. in their paper suggested the further investigation
of the relative importance between the aqueous and dietary
exposure of metals to fish in order to deeper understand the
processes of metal transfer into the fish body [30]. In agree-
ment with this conclusion in our last study we investigated the
Iron and Manganese Retention of Juvenile Zebrafish (Danio rerio) Exposed to Contaminated Dietary... 739
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
effect of waterborne Fe and Mn on the tissue of indicator fish,
while in present work the dietary Fe and Mn is tested. Trophic
transfer factor can be used for the latter purpose to express the
bioconcentration from zooplankton to fish [53]. The results of
TTF are more homogenous compared to BCF proving less
difference between the applied concentration of dietary Fe
and Mn. The values suggest that transfer definitely occurs
from zooplankton to fish for the studied metals yet in a lower
extent than directly from water to fish, as described in our
previous study [28]. According to the results of Zhu et al.
D. rerio could also accumulate nTiO
2
by aqueous exposure
with higher bioaccumulation factors compared to the dietary
intake.
Biomagnification is not observed in present study along the
water to D. pulex to D. rerio trophic route. This finding meets
previously published research data stating that BCF and TTF
usually increase along the given aquatic food chain right until
the level of fish where it starts to decrease significantly.
Pianpian et al. concluded in their meta-analysis that the aver-
age MeHg bioaccumulation from water to either seston or
zooplankton was over 6 order of magnitude greater than the
biomagnification from zooplankton to preyfish [66]. Mathews
et al. studied the retention ofMeHg, Cd, and Po in an estuarine
food chain and found a greater biomagnification at the trophic
step of D. pulex feeding on phytoplankton [67]. This phenom-
enon is commonly explained by the higher availability of tox-
ic elements to organisms at lower trophic levels [68]. Our
study further confirms Oweson and Hernroth finding that
Mn tends to accumulate strongly in aquatic organisms situated
lower in the food web [69].
It was also long found that aquatic vertebrates as fish can
efficiently excrete ingested heavy metals, especially the no-
essential and toxic ones [70]. Increased BCF factor for Fe and
Mn was observed from water to fish in our previous experi-
ment compared to the current one, where only in the absorp-
tion trough the respiratory and dermal surfaces was considered
[28]. It further indicates that retention of waterborne Fe and
Mn is stronger compared to the dietary one; however, the
accumulation extent of elements vary among the metals and
metal species showing very different behavioral patterns. For
example, the main route of Cd accumulation in marine envi-
ronment is considered to be the dietary exposure as well as
most of the other metals that have been studied in the past
decades [71]. For freshwater ecosystems Cd was found to
enrich strongly in the trophic chain, while no enrichment ten-
dency of Cu was observed in the same survey [72]. The accu-
mulation tendency of Mn was investigated by Niemiec et al. in
a field study resulting in much higher BCF values from water
to Ciprinus carpio in the analyzed ecosystem than other
routes.
It is also proven by several field studies and laboratory
experiments that toxic elements are accumulated in certain
organs contributing to a small extent to the total weight of
the fish body explaining the lower level of retention results.
This finding is confirmed for waterborne Fe and Mn in ourlast
experiment where both were absorbed in the highest level in
the liver of Common carp juveniles [28]. Metals stored in the
form of granules usually have lower bioavailability to the
predator organisms [73].
The so-called grow dilution may also take part inthe small-
er TTF values from zooplankton to D. rerio compared to the
BCF data given for water to D. pulex. This pseudo-elimination
process occurs due to the increase of fish tissue during the
experimental time and results in a lower determined BCF
values even more under laboratory circumstances [59,74].
Conclusion
Present work indicates the importance of model experiments to
gain more detailed information regarding the accumulation ten-
dency of metals, not limited solely to the non-essential ones. It
highlights that laboratory circumstances provide the possibility
to investigate only one absorption pathway at a time and to
selectively exclude the others. Iron and manganese are essential
elements to a certain level above which both have negative
influence on aquatic organisms. Our work proves that the main
step from the point of bioconcentrationoccursinthelowerpart
of the trophic system since higher accumulation tendency was
observed in the first stage of the investigated food web. The
results demonstrate that concentrations of Fe and Mn well
above the environmentally accepted levels still do not cause
mortality neither biomagnification in the studied organisms;
however, retention in the growth performance of D. rerio was
observed. The lack of acute toxicity thus does not translate into
no negative effect. It is mentioned in literature that damage to
chromosome levels and changes in locomotor and exploratory
behavior is resulted in long-term exposures. Further model
studies are therefore recommended to investigate the dietary
Fe and Mn absorption in the different organs and tissues of fish
to supplement research approaches determining the biochemi-
cal adverse effects, such as oxidative stress.
Acknowledgments We acknowledge the Agilent Technologies, Inc. and
the Novo-Lab Ltd. (Hungary) for providing the MP-AES 4200 and the
ICP-OES 5100 instruments for the elemental analysis.
Funding Information Open access funding provided by University of
Debrecen (DE). The research was supported by the EU and co-financed
by the European Regional Development Fund under the project GINOP-
2.3.2-15-2016-00008. Petra Herman was supported through the New
National Excellence Program of the Ministry of Human Capacities.
Compliance with Ethical Standards
Conflict of Interest The authors declare that they have no conflict of
interest.
Herman et al.
740
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Ethical Approval All experiments and procedures were performed in
compliance with relevant laws and institutional guidelines, and the ap-
propriate institutional committee have approved them.
Open Access This article is licensed under a Creative Commons
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References
1. Dural M, Genc E, Sangun MK, Güner Ö (2011) Accumulation of
some heavy metals in Hysterothylacium aduncum (Nematoda) and
its host sea bream, Sparus aurata (Sparidae) from North-Eastern
Mediterranean Sea (Iskenderun Bay). Environ Monit Assess 174:
147–155. https://doi.org/10.1007/s10661-010-1445-0
2. Frémion F, Bordas F, Mourier B, Lenain JF, Kestens T, Courtin-
Nomade A (2016) Influence of dams on sediment continuity: a
study case of a natural metallic contamination. Sci Total Environ
547:282–294. https://doi.org/10.1016/j.scitotenv.2016.01.023
3. Gheorghe S, Stoica C, Vasile GG, et al (2017) Metals toxic effects
in aquatic ecosystems: modulators of water quality. In: Tutu H (ed)
Water quality. InTech
4. Hasan MR, Khan MZH, Khan M, Aktar S, Rahman M, Hossain F,
Hasan ASMM (2016) Heavy metals distribution and contamination
in surface water of the Bay of Bengal coast. Cogent Environ Sci 2.
https://doi.org/10.1080/23311843.2016.1140001
5. Varol M, Şen B (2012) Assessment of nutrient and heavy metal
contamination in surface water and sediments of the upper Tigris
River, Turkey. CATENA 92:1–10. https://doi.org/10.1016/j.
catena.2011.11.011
6. Yılmaz AB, Yanar A, Alkan EN (2017) Review of heavy metal
accumulation on aquatic environment in Northern East
Mediterranean Sea part I: some essential metals. Rev Environ
Health 32:119–163. https://doi.org/10.1515/reveh-2016-0065
7. Zuur AF, Ieno EN, Smith GM (2007) Principal component analysis
and redundancy analysis. In: Analysing ecological data. Springer
New York, New York, NY, pp 193–224
8. Bonnail E, Sarmiento AM, DelValls TA et al (2016) Assessment of
metal contamination, bioavailability, toxicity and bioaccumulation
in extreme metallic environments (Iberian Pyrite Belt) using
Corbicula fluminea. Sci Total Environ 544:1031–1044. https://
doi.org/10.1016/j.scitotenv.2015.11.131
9. Griscom SB, Fisher NS (2004) Bioavailability of sediment-bound
metals to marine bivalve molluscs: An overview. Estuaries 27:826–
838. https://doi.org/10.1007/BF02912044
10. Kwak JI, Cui R, Nam S-H, Kim SW, Chae Y, An YJ (2016)
Multispecies toxicity test for silver nanoparticles to derive hazard-
ous concentration based on species sensitivity distribution for the
protection of aquatic ecosystems. Nanotoxicology 10:521–530.
https://doi.org/10.3109/17435390.2015.1090028
11. Zhou Q, Zhang J, Fu J, Shi J, Jiang G (2008) Biomonitoring: an
appealing tool for assessment of metal pollution in the aquatic eco-
system. Anal Chim Acta 606:135–150. https://doi.org/10.1016/j.
aca.2007.11.018
12. Cui R, Kwak JI, An Y-J (2018) Comparative study ofthe sensitivity
of Daphnia galeata and Daphnia magna to heavy metals.
Ecotoxicol Environ Saf 162:63–70. https://doi.org/10.1016/j.
ecoenv.2018.06.054
13. Lari E, Gauthier P, Mohaddes E, Pyle GG (2017) Interactive tox-
icity of Ni, Zn, Cu, and Cd on Daphnia magna at lethal and sub-
lethal concentrations. J Hazard Mater 334:21–28. https://doi.org/10.
1016/j.jhazmat.2017.03.060
14. Manakul P, Peerakietkhajorn S, Matsuura T, Kato Y, Watanabe H
(2017) Effects of symbiotic bacteria on chemical sensitivity of
Daphnia magna. Mar Environ Res 128:70–75. https://doi.org/10.
1016/j.marenvres.2017.03.001
15. Bownik A (2017) Daphnia swimming behaviour as a biomarker in
toxicity assessment:a review. Sci Total Environ 601–602:194–205.
https://doi.org/10.1016/j.scitotenv.2017.05.199
16. Cerveny D, Turek J, Grabic R, Golovko O, Koba O, Fedorova G,
Grabicova K, Zlabek V, Randak T (2016) Young-of-the-year fish
as a prospective bioindicator for aquatic environmental contamina-
tion monitoring. Water Res 103:334–342. https://doi.org/10.1016/j.
watres.2016.07.046
17. Habib MR, Mohamed AH, Osman GY, Mossalem HS, Sharaf el-
Din AT, Croll RP (2016) Biomphalaria alexandrina as a
bioindicator of metal toxicity. Chemosphere 157:97–106. https://
doi.org/10.1016/j.chemosphere.2016.05.012
18. Łuczyńska J, Paszczyk B, Łuczyński MJ (2018) Fish as a
bioindicator of heavy metals pollution in aquatic ecosystem of
Pluszne Lake, Poland, and risk assessment for consumer’s health.
Ecotoxicol Environ Saf 153:60–67. https://doi.org/10.1016/j.
ecoenv.2018.01.057
19. Bhuvaneshwari M, Iswarya V, Vishnu S, Chandrasekaran N,
Mukherjee A (2018) Dietary transfer of zinc oxide particles from
algae (Scenedesmus obliquus)todaphnia(Ceriodaphnia dubia).
Environ Res 164:395–404. https://doi.org/10.1016/j.envres.2018.
03.015
20. Nguyen VT, Satoh S, Haga Y, Fushimi H, Kotani T (2008) Effect
of zinc and manganese supplementation in Artemia on growth and
vertebral deformity in red sea bream (Pagrus major) larvae.
Aquaculture 285:184–192. https://doi.org/10.1016/j.aquaculture.
2008.08.030
21. Skjolding LM, Winther-Nielsen M, Baun A (2014) Trophic transfer
of differently functionalized zinc oxide nanoparticles from crusta-
ceans (Daphnia magna) to zebrafish (Danio rerio). Aquat Toxicol
157:101–108. https://doi.org/10.1016/j.aquatox.2014.10.005
22. Zhu X, Wang J, Zhang X, Chang Y, Chen Y (2010) Trophic trans-
fer of TiO
2
nanoparticles from daphnia to zebrafish in a simplified
freshwater food chain. Chemosphere 79:928–933. https://doi.org/
10.1016/j.chemosphere.2010.03.022
23. Parmar TK, Rawtani D, Agrawal YK (2016) Bioindicators: the
natural indicator of environmental pollution. Front Life Sci 9:
110–118. https://doi.org/10.1080/21553769.2016.1162753
24. Araujo GS, Pavlaki MD, Soares AMVM, Abessa DMS, Loureiro S
(2019) Bioaccumulation and morphological traits in a multi-
generation test with two Daphnia species exposed to lead.
Chemosphere 219:636–644. https://doi.org/10.1016/j.
chemosphere.2018.12.049
25. Lu K, Qiao R, An H, Zhang Y (2018) Influence of microplastics on
the accumulation and chronic toxic effects of cadmium in zebrafish
(Danio rerio). Chemosphere 202:514–520. https://doi.org/10.1016/
j.chemosphere.2018.03.145
26. Zhang J, Hamza I (2018) Zebrafish as a model system to delineate
the role of heme and iron metabolism during erythropoiesis. Mol
Genet Metab 128:204–212. https://doi.org/10.1016/j.ymgme.2018.
12.007
27. Fernández I, Gavaia PJ, Laizé V, Cancela ML (2018) Fish as a
model to assess chemical toxicity in bone. Aquat Toxicol 194:
208–226. https://doi.org/10.1016/j.aquatox.2017.11.015
Iron and Manganese Retention of Juvenile Zebrafish (Danio rerio) Exposed to Contaminated Dietary... 741
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
28. Harangi S, Baranyai E, Fehér M, Tóth CN, Herman P, Stündl L,
Fábián I, Tóthmérész B, Simon E (2017) Accumulation of metals in
juvenile carp (Cyprinus carpio) exposed to sublethal levels of iron
and manganese: survival, body weight and tissue. Biol Trace Elem
Res 177:187–195. https://doi.org/10.1007/s12011-016-0854-5
29. Koivisto S (1995) Is Daphnia magnaan ecologically representative
zooplankton species in toxicity tests? Environ Pollut 90:263–267.
https://doi.org/10.1016/0269-7491(95)00029-Q
30. Qu R-J, Wang X-H, Feng M-B, Li Y, Liu HX, Wang LS, Wang ZY
(2013) The toxicity of cadmium to three aquatic organisms
(Photobacterium phosphoreum,Daphnia magna and Carassius
auratus) under different pH levels. Ecotoxicol Environ Saf 95:83–
90. https://doi.org/10.1016/j.ecoenv.2013.05.020
31. Tan C, Wang W-X (2014) Modification of metal bioaccumulation
and toxicity in Daphnia magna by titanium dioxide nanoparticles.
Environ Pollut 186:36–42. https://doi.org/10.1016/j.envpol.2013.
11.015
32. Cardwell RD, DeForest DK, Brix KV, Adams WJ (2013) Do cd,
cu, Ni, Pb, and Zn biomagnify in aquatic ecosystems? In: Whitacre
DM (ed) Reviews of environmental contamination and toxicology,
vol 226. Springer New York, New York, NY, pp 101–122
33. Rubio-Franchini I, López-Hernández M, Ramos-Espinosa MG,
Rico-Martínez R (2016) Bioaccumulation of metals arsenic, cadmi-
um, and lead in zooplankton and fishes from the Tula River
Watershed, Mexico. Water Air Soil Pollut 227:5. https://doi.org/
10.1007/s11270-015-2702-1
34. Oberholster PJ, Myburgh JG, Ashton PJ, Coetzee JJ, Botha AM
(2012) Bioaccumulation of aluminium and iron in the food chain of
Lake Loskop, South Africa. Ecotoxicol Environ Saf 75:134–141.
https://doi.org/10.1016/j.ecoenv.2011.08.018
35. Chandra Sekhar K, Chary NS, Kamala CT, Suman Raj DS,
Sreenivasa Rao A (2004) Fractionation studies and bioaccumula-
tion of sediment-bound heavy metals in Kolleru lake by edible fish.
Environ Int 29:1001–1008. https://doi.org/10.1016/S0160-
4120(03)00094-1
36. Rajeshkumar S, Li X (2018) Bioaccumulation of heavy metals in
fish species from the Meiliang Bay, Taihu Lake, China. Toxicol
Rep 5:288–295. https://doi.org/10.1016/j.toxrep.2018.01.007
37. Jitar O, Teodosiu C, Oros A, Plavan G, Nicoara M (2015)
Bioaccumulation of heavy metals in marine organisms from the
Romanian sector of the Black Sea. New Biotechnol 32:369–378.
https://doi.org/10.1016/j.nbt.2014.11.004
38. Mathews T, Fisher N (2008) Trophic transfer of seven trace metals
in a four-step marine food chain. Mar Ecol-Prog Ser - MARECOL-
PROGR SER 367:23–33. https://doi.org/10.3354/meps07536
39. Hwang D-W, Kim S-S, Kim S-G, Kim DS, Kim TH (2017)
Erratum to: Concentrations of heavy metals in marine wild fishes
captured from the Southern Sea of Korea and associated health risk
assessments. Ocean Sci J 52:467–467. https://doi.org/10.1007/
s12601-017-0048-x
40. Bosch AC, O’Neill B, Sigge GO et al (2016) Heavy metals in
marine fish meat and consumer health: a review: heavy metals in
marine fish meat. J Sci Food Agric 96:32–48. https://doi.org/10.
1002/jsfa.7360
41. Feng X, Meng B, Yan H, Fu X, Yao H, Shang L, Feng X, Meng B,
Yan H, Fu X, Yao H, Shang L (2018) Bioaccumulation of mercury
in aquatic food chains. In: Biogeochemical cycle of mercury in
reservoir systems in Wujiang River Basin. Southwest China.
Springer Singapore, Singapore, pp 339–389
42. Lavoie RA, Jardine TD, Chumchal MM, Kidd KA, Campbell LM
(2013) Biomagnification of mercury in aquatic food webs: a world-
wide meta-analysis. Environ Sci Technol 47:13385–13394. https://
doi.org/10.1021/es403103t
43. Espejo W, Padilha J de A, Kidd KA, et al (2018) Trophic transfer of
cadmium in marine food webs from Western Chilean Patagonia and
Antarctica. Mar Pollut Bull 137:246–251. https://doi.org/10.1016/j.
marpolbul.2018.10.022
44. Leszczyńska K (1997) The accumulation of Cd, Pb and Cu in the
aquatic food chains in three lakes differing in the trophic conditions.
SIL Proc 1922-2010 26:517–519. https://doi.org/10.1080/
03680770.1995.11900769
45. World Health Organization (ed) (2011) Guidelines for drinking-
water quality, 4th edn. World Health Organization, Geneva
46. Ekström SM, Regnell O, Reader HE, Nilsson PA, Löfgren S,
Kritzberg ES (2016) Increasing concentrations of iron in surface
waters as a consequence of reducing conditions in the catchment
area. J Geophys Res Biogeosci 121:479–493. https://doi.org/10.
1002/2015jg003141
47. Balcu I, Amalia C, Segneanu A-E, Oana R (2011) Combined
microwave-acid pretreatment of the biomass. In: Shaukat S (ed)
Progress in biomass and bioenergy production. InTech
48. Balogh Z, Harangi S,Kundrát JT, Gyulai I, Tóthmérész B, Simon E
(2016) Effects of anthropogenic activities on the elemental concen-
tration in surface sediment of oxbows. Water Air Soil Pollut 227.
https://doi.org/10.1007/s11270-015-2714-x
49. Balogh Z, Harangi S, Gyulai I, Braun M, Hubay K, Tóthmérész B,
Simon E (2017) Exploring river pollution based on sediment anal-
ysis in the Upper Tisza region (Hungary). Environ Sci Pollut Res
24:4851–4859. https://doi.org/10.1007/s11356-016-8225-5
50. Ódor L, Horváth I, Fügedi U (1997) Low-density geochemical
mapping in Hungary. J Geochem Explor 60:55–66. https://doi.
org/10.1016/S0375-6742(97)00025-3
51. AVMA Guidelines for the Euthanasia of Animals (2013) Edition.
https://norecopa.no/textbase/avma-guidelines-for-the-euthanasia-
of-animals-2013-edition. Accessed 23 Apr 2019
52. Fehér M, Baranyai E, Simon E, Bársony P, Szűcs I, Posta J, Stündl
L (2013) The interactive effect of cobalt enrichment in Artemia on
the survival and larval growth of barramundi, Lates calcarifer.
Aquaculture 414–415:92–99. https://doi.org/10.1016/j.
aquaculture.2013.07.031
53. Handbook of property estimation methods for chemicals: environ-
mental health sciences. In: CRC Press. https://www.crcpress.com/
Handbook-of-Property-Estimation-Methods-for-Chemicals-
Environmental-Health/Mackay-Boethling/p/book/
9781566704564. Accessed 23 Apr 2019
54. (2007) Principal component analysis and redundancy analysis. In:
Analysing ecological data. Springer New York, New York, NY, pp
193–224
55. Tuzuki BLL, Delunardo FAC, Ribeiro LN et al (2017) Effects of
manganese on fat snook Centropomus parallelus (Carangaria:
Centropomidae) exposed to different temperatures. Neotropical
Ichthyol 15. https://doi.org/10.1590/1982-0224-20170054
56. Peters A, Lofts S, Merrington G, Brown B, Stubblefield W, Harlow
K (2011) Development of biotic ligand models for chronic manga-
nese toxicity to fish, invertebrates, and algae. Environ Toxicol
Chem 30:2407–2415. https://doi.org/10.1002/etc.643
57. Dolci GS, Dias VT, Roversi K, Roversi K, Pase CS, Segat HJ,
Teixeira AM, Benvegnú DM, Trevizol F, Barcelos RCS, Riffel
APK, Nunes MAG, Dressler VL, Flores EMM, Baldisserotto B,
Bürger ME (2013) Moderate hypoxia is able to minimize the
manganese-induced toxicity in tissues of silver catfish (Rhamdia
quelen). Ecotoxicol Environ Saf 91:103–109. https://doi.org/10.
1016/j.ecoenv.2013.01.013
58. Gabriel D, Riffel APK, Finamor IA, Saccol EMH, Ourique GM,
Goulart LO, Kochhann D, Cunha MA, Garcia LO, Pavanato MA,
Val AL, Baldisserotto B, Llesuy SF (2013) Effects of subchronic
manganese chloride exposure on Tambaqui (Colossoma
macropomum) tissues: oxidative stress and antioxidant defenses.
Arch Environ Contam Toxicol 64:659–667. https://doi.org/10.
1007/s00244-012-9854-4
Herman et al.
742
Content courtesy of Springer Nature, terms of use apply. Rights reserved.
59. Adolfsson-Erici M, Åkerman G, McLachlan MS (2012) Measuring
bioconcentration factors in fish using exposure to multiple
chemicals and internal benchmarking to correct for growth dilution.
Environ Toxicol Chem 31:1853–1860. https://doi.org/10.1002/etc.
1897
60. Vieira MC, Torronteras R, Córdoba F, Canalejo A (2012) Acute
toxicity of manganese in goldfish Carassius auratus is associated
with oxidative stress and organ specific antioxidant responses.
Ecotoxicol Environ Saf 78:212–217. https://doi.org/10.1016/j.
ecoenv.2011.11.015
61. Marins K, Lazzarotto LMV, Boschetti G, Bertoncello KT, Sachett
A, Schindler MSZ, Chitolina R, Regginato A, Zanatta AP, Siebel
AM, Magro JD, Zanatta L (2019) Iron and manganese present in
underground water promote biochemical, genotoxic, and behavior-
al alterations in zebrafish (Danio rerio). Environ Sci Pollut Res 26:
23555–23570. https://doi.org/10.1007/s11356-019-05621-0
62. Tu H, Fan C, Chen X, Liu J, Wang B, Huang Z, Zhang Y, Meng X,
Zou F (2017) Effects of cadmium, manganese, and lead on loco-
motor activity and neurexin 2a expression in zebrafish: effects of
metals on neurexin 2a expression in zebrafish. Environ Toxicol
Chem 36:2147–2154. https://doi.org/10.1002/etc.3748
63. Altenhofen S, Wiprich MT, Nery LR, Leite CE, Vianna MRMR,
Bonan CD (2017) Manganese (II) chloride alters behavioral and
neurochemical parameters in larvae and adult zebrafish. Aquat
Toxicol 182:172–183. https://doi.org/10.1016/j.aquatox.2016.11.
013
64. Voigt CL, da Silva CP, Doria HB, Randi MAF, de Oliveira Ribeiro
CA, de Campos SX (2015) Bioconcentration and bioaccumulation
of metal in freshwater Neotropical fish Geophagus brasiliensis.
Environ Sci Pollut Res 22:8242–8252. https://doi.org/10.1007/
s11356-014-3967-4
65. Vuori K-M (1995) Direct and indirect effects of iron on river eco-
systems. Ann Zool Fenn 32:317–329
66. Wu P, Kainz MJ, Bravo AG, Åkerblom S, Sonesten L, Bishop K
(2019) The importance of bioconcentration into the pelagic food
web base for methylmercury biomagnification: a meta-analysis. Sci
Total Environ 646:357–367. https://doi.org/10.1016/j.scitotenv.
2018.07.328
67. Mathews T, Fisher NS (2008) Evaluating the trophic transfer of
cadmium, polonium, andmethylmercury in an estuarine food chain.
Environ Toxicol Chem 27:1093–1101. https://doi.org/10.1897/07-
318.1
68. Dallinger R, Prosi F, Segner H, Back H (1987) Contaminated food
and uptake of heavy metals by fish: a review and a proposal for
further research. Oecologia 73:91–98. https://doi.org/10.1007/
BF00376982
69. Oweson C, Hernroth B (2009) A comparative study on the influ-
ence of manganese on the bactericidal response of marine inverte-
brates. Fish Shellfish Immunol 27:500–507. https://doi.org/10.
1016/j.fsi.2009.07.001
70. Kawasaki LY, Tarifeño-Silva E, Yu DP, Gordon MS, Chapman DJ
(1982) Aquacultural approaches to recycling of dissolved nutrients
in secondarily treated domestic wastewaters—I Nutrient uptake and
release by artificial food chains. Water Res 16:37–49. https://doi.
org/10.1016/0043-1354(82)90051-3
71. Wang W (2002) Interactions of trace metals and different marine
food chains. Mar Ecol Prog Ser 243:295–309. https://doi.org/10.
3354/meps243295
72. Croteau M-N, Luoma SN, Stewart AR (2005) Trophic transfer of
metals along freshwater food webs: evidence of cadmium
biomagnification in nature. Limnol Oceanogr 50:1511–1519.
https://doi.org/10.4319/lo.2005.50.5.1511
73. Wg W, Gr L (1997) Bioavailability of biologically sequestered
cadmium and the implications of metal detoxification. Mar Ecol
Prog Ser 147:149–157. https://doi.org/10.3354/meps147149
74. Arnot JA, Gobas FA (2006) A review of bioconcentration factor
(BCF) and bioaccumulation factor (BAF) assessments for organic
chemicals in aquatic organisms. Environ Rev 14:257–297. https://
doi.org/10.1139/a06-005
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