Egyptian Journal of Aquatic Research 45 (2019) 337–343

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Egyptian Journal of Aquatic Research

journal homepage: www.sciencedirect.com/locate/ejar

The effect of cadmium exposure on the cytoskeleton and morphology of

the gill chloride cells in juvenile mosquito fish (Gambusia affinis)
Moh. Awaludin Adam a, Maftuch Maftuch b, Yuni Kilawati b, Yenny Risjani b,⇑
a
Faculty of Sciences and Technology, University of Ibrahimy, Situbondo, Indonesia
b
Faculty of Fisheries and Marine Sciences, Universitas Brawijaya, Indonesia

a r t i c l e i n f o a b s t r a c t

Article history: One of the wide spread aquatic organisms in the rivers of Indonesia is the mosquito fish (Gambusia affinis
Received 30 July 2018 (Baird and Girard, 1853)). Mosquito fish are invasive species that can adapt to environmental changes,
Revised 16 November 2019 also they act as bioindicator species for polluted environments. The study aims to determine the effects
Accepted 30 November 2019
of the exposure to cadmium on the cytoskeleton and morphology of the gill’s Chloride Cells (CCs) in the
Available online 20 December 2019
mosquito fish. In cadmium treated mosquito fish (0.03 mg/L for 28 days), the immuno-fluorescent light
microscope showed strong absorption of actin stain and strong primary MT antibody uptake in CC and
Keywords:
apical epithelia. Scanning electron microscopy observations showed a change in microfilament organized
Cadmium
Chloride cells
at the CC’s apex, with the appearance of some actin filament aggregates. Higher cadmium concentrations
Cytoskeleton (0.03 and 0.015 mg/L) did not alter such reorganization. Microtubules weren’t significantly affected by
Gambusia affinis similar exposures. SEM analysis showed that cadmium exposure induced a significant increase in CC’s
Microfilaments surface area. After 28 days, the density of CC also increased. It was observed that there is an increase
of CC’s surface area, although the CC density didn’t increase significantly. The results of this study con-
firmed the cytotoxic effects of cadmium on mosquito fish.
Ó 2019 National Institute of Oceanography and Fisheries. Hosting by Elsevier B.V. This is an open access
article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

Introduction
The contamination of the aquatic environment by heavy metals
has caused much anxiety especially regarding its effects on the aqua-
tic organisms (Adam et al., 2018). One of the wide-spread aquatic
organisms in the river streams of Indonesia is the mosquito fish
(Gambusia affinis) (Baird and Girard, 1853). Gambusia affinis has
become a biological control agent for mosquitoes (Carlson et al.,
2015). It is considered to be as effective as the native predators of
mosquitoes (Global Invasive Species Database GISD, 2015). These
fish live in many water bodies. During the growing period of the
fish’s lifecycle, complex mechanisms develop that allow fish to adapt
to changes in the environment (El-Greisy and El-Gamal, 2015).
Mosquito fish is an invasive species that can adapt to environ-
mental changes (Lamatsch et al., 2015). Beside their function as a
vital breathing organ, the gills of mosquito fish, act as the principal
organ for ion regulation, as well as functioning as bio-indicator for
polluted environments (Hemmadi, 2016). The gill is a multipur-
Peer review under responsibility of National Institute of Oceanography and
Fisheries.
⇑ Corresponding author.
E-mail addresses: ar.adam87@yahoo.com (M.A. Adam), risjani@ub.ac.id
(Y. Risjani).
pose organ that, in addition to providing for aquatic gas exchange
and excretion of nitrogenous wastes, plays dominant roles in
osmotic, ionic, and acid-base regulation (Evans et al., 2005). These
cells are widely studied, considering their importance in vital func-
tions of the gill. The cells proliferate when exposed to unfavorable
environments (Pereira and Caetano, 2009).
During the early years, G. affinis not only faced osmoregulatory
problems but often also pollution. The latter is highly cytotoxic
because it interferes with other cation functions, as influencing
nutrient stability and bioavailability (Hemmadi, 2016), and acidity
(pH) especially with regard to calcium (Flik and Verbost, 1993).
The impacts on fish physiology, and even on some gill function
of freshwater fish, such as tilapia, as well as the effect of heavy
metal concentrations in the gills of Oreochromis mossambicus are
well documented (Bonga et al., 1990). The Morphology of CC in
various species indicates the adaptive changes in the number and
volume of CCs that occur when the ions balance is disrupted by
factors such as water salinity variation (Bonga and Van Der Meij,
1989), acidification of water (Pisam et al., 1995), or hormonal
changes (Perry and Wood, 1985). It’s also well documented that
during the natural aging process, the gill epithelium goes through
marked modification in the CC’s number (Lin and Sung, 2003).
Several studies have shown that the regulation of the cell vol-
ume in is osmotic conditions, at least partially, depends on an

https://doi.org/10.1016/j.ejar.2019.11.011
1687-4285/Ó 2019 National Institute of Oceanography and Fisheries. Hosting by Elsevier B.V.
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
338 M.A. Adam et al. / Egyptian Journal of Aquatic Research 45 (2019) 337–343
intact cytoskeleton (Prat et al., 1996). Furthermore, the phe-
nomenon of the volume regulation coincides with the reorganiza-
tion of a large-scale actin filaments. The structural modification
may depend on increased osmotic-influenced intra cellular cal-
cium concentration (Guilak et al., 2002). In both freshwater and
seawater fish, the underlying Ca2+ active transport uptakes
transepithelial Ca2+ (Verbost et al., 1994). Cadmium can disrupt
the process of transporting calcium, and induces cytosolic calcium
reactions in CC (Flik et al., 1993).
Cadmium (Cd) is a heavy metal that can pollute aquatic ecosys-
tems (Annabi et al., 2011). It is accumulative in aquatic biota and
causes abnormalities in fish (Asagba et al., 2008). The defects that
occur are tissue damage (Hertika et al., 2019), osmoregulation dis-
orders (Soegianto et al., 2008) and changes in habits (Scott, 2003).
Accumulation of cadmium in the fish body occurs in gill organs
(Galvez et al., 2006), liver (Min et al., 2016) and kidney (Vesey,
2010). Detection of the accumulation of cadmium (Adam et al.,
2019b) can be a metallothionein (MT) biomarker (Van Heerden
et al., 2006). MT expression describes the level of uptake of cad-
mium (Yang et al., 2017) and is considered as a diagnostic tool in
fish (Bakiu et al., 2013).
The study aims to determine the effects of exposure to cad-
mium on the cytoskeleton and morphology of the CC of the gills
in young Gambusia fish.
Materials and methods
Animal experiments
The young mosquito fish (Gambusia affinis) were collected from
the Freshwater Aquaculture Installation (IBAT), Punten, Batu City,
East Java. The characteristics of the collected sample showed
healthy fish, sexually immature, weighing between 1 and 2 g with
a length of 3–5 cm. Young gambusia fish were brought to the labo-
ratory of University of Brawijaya (UB) Fish Reproduction and main-
tained for two weeks while being placed in an aquarium, which was
filled with water (4 L). Each aquarium was stocked with ten fish.
Cadmium exposure
One hundred and eighty fish G. affinis were divided into 18
aquaria; 10 fish per aquarium. Each aquarium contains 2 L of de-
chlorinated static water. Three replications represented each con-
centration of cadmium. Each group of fish was exposed to cad-
mium concentration of LC50 carried out for 28 days as follows:
Group A: 0:03 mg/L of water (100% of the LC50); Group B:
0.015 mg/L of water (75% of the LC50); Group C: 0.0115 mg/L of
water (50% of the LC50) and group D: 0.0075 mg/L of water (25%
of the LC50), group E: 0.0035 mg/L of water (12.5% of the LC50)
and group K (control) stored in chlorinated water containing 0.0%
cadmium. More than 30% of exposure was lost due to volatilization
(Ibrahem, 2011). The water in the treatments and the control was
exchanged every day. The fish were fed silkworms (Tubifex sp). The
fecal waste were siphoned every day.
Water quality observation
The quality of water in the laboratory was kept stable, to min-
imize the influence of the environment. The water quality observa-
tions are held in Table 1.
Preparation and histological observation gills
Gills of young gambusia fish exposed to Cd were fixed using 10%
phosphate-buffered formalin (NBF) for 24–48 h (Mumford et al.,
Table 1
Water Quality Observations.
Water Quality Parameter Value
Temperature 23 ± 4 °C
Dissolved Oxygen (DO) 4,6 ± 1,4 mg/L
pH 6,0 ± 1,2
Note: Average water quality results during the observation.
2017). Fixation activities were conducted at the Pathology Labora-
tory, Faculty of Medicine, University of Brawijaya Malang. This was
followed by tissue processing to maintain cell and tissue condi-
tions. Blocks were formed from the tissue embedding process.
The tissue is cut using a microtome. The section thickness is 3–
5 mm with the cutting tape stretched in a stable temperature of
40° C. Glass slides are used to take ribbons in the water bath. Then,
they were air dried for one hour. Afterwards, the glass slides were
arranged in a staining box and stained with hematoxylin-eosin
(HE) (Pereira and Caetano, 2009). At the end of the staining, a cover
slip was mounted on the tissue specimen on the slide. Preparations
were examined with an Olympus microscope and documented
using the dot slide method. The data were analyzed descriptively
by looking at changes in tissue by exposure to Cd.
Fluorescence staining with confocal laser scanning microscopy (CLSM)
After dissection, the staining method the Central Laboratory of
Biological Sciences (LSIH) standard, University of Brawijaya. Prepa-
rations were made by the modification method (Walker, 2014).
Paraffin sectioning were kept in oven at 40 °C overnight. Then
immersed in xylol two times for 10 min each. Followed by concen-
tration of ethanol immersion (1 and 2, 90% and 70%, respectively
for 5 min. Then, they were soaked in Phosphate Buffered Saline
with Tween-20 (PBST) three times, for 5 minutes each. Afterwards,
they were dipped in citrate buffer ten mM at pH 6 for 15 min at a
temperature of 120 °C. The preparations and the containers are
removed from the oven and left to settle for ±10 min, then soaked
again in PBST 3 times, for 5 min each. Then blocks were prepared
using 2% Bovine Serum Albumin (BSA) in PBST at room tempera-
ture for ±60 min, and washed by PBST 3 times, 8 min each. Then
blocked using primary antibody in BSA 2% (1:1000) for ±60 min,
washed with PBST 3 times, 8 min each. This was followed by sec-
ondary antibody dilution in 2% BSA (1:1500) for ±60 min. Again
slides were washed with PBST 3 times, 8 min each, and finally
drained and cleaned with a tissue then placed in 10% glycerol.
Scanning electron microscopy (SEM)
Preparations were made by the modification method (Devos
et al., 1998). After dissection, the paraffin was removed from sec-
tions and they were thinly cut to 2–3 mm thick and immediately
immersed with 2% glutaraldehyde in 0.05 M osmium tetraoxide
(pH: 7.4; fixative osmotic pressure: 310 mosmol l-1) for 90 min.
Then they were set in room temperature for 4  10 min each in
buffer osmium tetraoxide 0.15 M. Afterwards, they were left to
dry for 5 min then put in acetone solution (30, 50, 70, 90 and
100%) respectively for 5 min and dried at critical point (critical
point dryer, Balzers CPD 030). The sample was mounted on a silver
paint stub by maintaining the primary lamella parallel to the stub,
and then gold coated by the sputtering method (Balzers). Scanning
electron microscopy with Philips XL 20. For each experimental
condition, five fish were examined. Chloride cells were found on
the trailing edges of the first filaments in the interlamellar area
and photographed at 8000 magnification. At least four non-
 M.A. Adam et al. / Egyptian Journal of Aquatic Research 45 (2019) 337–343 339

contiguous areas were randomly selected from each fish for mor-
phometric analysis.
Image analysis
Average CC surface area (lm2) and surface CC density (cells per
mm2) were determined after the method (Perry and Wood, 1985)
by tracking CC perimeter using a morphometric software program
(ImageJ). Also, for each micrograph and from these measurable
parameters, the fractional surface area of the chloride cell is calcu-
lated on the surface percent of the epithelial unit.
Statistic analysis
All scanning data are presented as means of ±1 SEM. Student’s
unpaired t-test was used to analyze the data labeled in Fig. 4. A
two-way ANOVA was used to analyze all other scanning data with
the program Statistical Package for the Social Sciences (SPSS) version
16 (Bryman and Cramer, 2005). Five percent was included as fidu-
ciary significance limits in all cases.
Results
Morphology of gills
Compared with control fish, gill morphology of exposed cad-
mium fish was comparable to that of other endangered fish. The
secondary lamella is coated by epithelium which is a thick layer
of squamous cell, while the epithelial layer stratum covers the first
filaments. Epithelial gill cells are made of more substantial parts
such as pavement cells (PVC), some muscles cells and some chlo-
ride cells (CC). Cadmium, at treatments of D and E for a period of
3–4 weeks, did not change the gill morphology. However, in the
treatment of A, B, and C, changes were observed on some filaments,
e.g. removal of epithelium, rupture and cell hypertrophy in sec-
ondary lamellae. For more results, the changes are shown in Figs. 1
and 4.
The entire length of the primary and secondary lamella are
lined by epithelial cells. In some cases, having a cytoplasm slightly
colored with Hematoxylin (Fig. 1). Only a few cells located in the
secondary lamella (Fig. 1B) and in between (Fig. 1A–E) show the
dark granules throughout the cytoplasm. The granules are typical
of the method of Von Kossa and occur at acidic pH. This was con-
firmed by the cytoplasm of cells with little purple-stained Hema-
toxylin. These cells were identified as chloride cells.
Staining of cytoskeleton
In the gill preparations of both treatments and control, microfil-
amentous tissues of primary and secondary filaments were stained
using MT antibodies and showed homogeneous epithelial pigmen-
tation of the epithelial cells (Fig. 2).
The results of the cytoskeleton staining has already demon-
strated the absorption of primary and secondary antibodies with
rhodamines. The low rate of the uptake depends on the amount
of cadmium concentration expressed. Fig. 2A shows that the

Fig. 1. Morphology of Gambusia affinis gills showing the difference between control and treatment groups with exposure to Cd (Group A: 0:03 mg/L of water (100% of the
LC50); Group B: 0.015 mg/L of water (75% of the LC50); Group C: 0.0115 mg/L of water(50% of the LC50) and group D: 0.0075 mg/L of water (25% of the LC50), group E:
0.0035 mg/L of water (12.5% of the LC50) and group K (control) stored in chlorinated water containing 0.0% cadmium) at a magnification of 20 lm. All images show the
existence of CC that are stained by HE.
340 M.A. Adam et al. / Egyptian Journal of Aquatic Research 45 (2019) 337–343
Fig. 2. The morphology of the gills of Gambusia affinis exposed to Cd, observed by
Rhodamin-Fluorescent staining by absorption of the MT enzyme and observed by a
confocal microscope (CL-SM). At P > 0.05, the intensity of MT density shows that the
treatments of A, B and C are very different from those of D and E against K. Full data
are not shown.
results of dye uptake is higher because of greater cadmium concen-
trations exposure. Likewise with Fig. 2B, 2C, and 2D, indicating an
increasingly lower absorption rate. As for Fig. 2E and K, they show
the results of uptake of dyes that are similarly low.
Fig. 3. Morphological gambusia fish granules, (A) indicates fish gill microfilaments without treatment; (B) shows microfilament with Cd exposure treatments (high
concentration).
Fig. 4. Morphological cross-section of chloride cells in sample A (without treatment of Cd exposure) and sample B (Cd exposure treatment-high concentration) showing the
wide variety of surface and density variations in the CC & PVC; initially with narrow breadth of high density. After exposure, they have become distant and with an enlarged
breadth.
SEM image
The SEM microscope image representation of fish gills in control
and treated cadmium are shown in Fig. 3. When observed under
normal conditions, gillarch has afringe of approximately 243.4 to
291.8 lm in length while for treated fish, the gill arch has a length
of about 100.7 lm with blunt gill arch display (proliferation). Fig. 4
is showing normal conditions where the apex of CC appears flat or
slightly concave. These cells show irregular margins but not cov-
ered by adjacent PVCs. With the presence of cadmium, CC appears
more regular and is often assumed to be a polyhedric configura-
tion, but still found adjacent to PVCs. The gills offish with CC are
characterized by different apical membranes from neighboring
adjacent cells showing apical membrane microridges rather than
microvilli (Figs. 3 and 4). Our SEM observations made it impossible
to detect morphological differences between the surface of chlo-
ride cells located at the base of the secondary lamella, sometimes
called a-chloride cells (Pisam and Rambourg, 1991) and those
found in the interlamellar region (b-chloride cells). In some cells,
the apical ornament is typical, where as elsewhere (even in the
same interlayer region), the apical membrane appears smoother,
due to rare or absent microvilli. It does not depend on cadmium
high concentration in different treatments and in control.
SEM analysis
The variations of the density and surface area of CC in normal
gambusia fish gills and exposed to cadmium are shown in Fig. 4.
The reference value (denoted as day 0) illustrates how different
values for control fish (A) are given under the same acclimation
and handling conditions than cadmium-treated fish; cadmium
high concentration (B) is randomly tested throughout the trial per-
iod from day 0 to day 28. The average value of this control fish does
not indicate the heterogeneity of the variance. SEM micrograph
 M.A. Adam et al. / Egyptian Journal of Aquatic Research 45 (2019) 337–343
Fig. 5. EDAx results from SEM for control fish (A) and fish treated with cadmium exposure-high concentration (B). Full data are not shown.
analysis showed significant differences between the control and
the treated ones. The surface area and fractional areas increase
from the total area 99.1% after exposure to 147.8% of the fish values
of high to low concentration. No significant variations are observed
with different CC density. The curve in Fig. 5 shows two distinct
components, i.e. cell surface area and cell density.
The Edax results regarding the concentration of the fish in con-
trol showed a rate of 0.02 mg/L (threshold 0.03 mg/L). The means
are still below the threshold for control fish, where as the concen-
tration of Edax results for fish given exposure to cadmium
increased to 0.28 mg/L. Such exposure is dangerous to the life of
the organism, which causes damage to the morphological appear-
ance of fish gills with treatment.
Discussion
The results of the present study evaluate the Cd impacts on the
gills of young Gambusia fish in terms of histopathological and his-
tochemical analysis. The evaluation showed the damage of gill
morphology and increase of the number and form of chloride cells.
The results of similar studies (Bonga et al., 1990), in freshwater fish
Tilapia (Oreochromis mossambicus), showed the cell death by necro-
sis, that only occurred transiently when the reduction of water pH
was acute (48 h) along with an increase in the number of chloride
cells. Other studies (Saibu et al., 2018), indicated that an acute
exposure to metals (mix Cu, Cd, Zn), resulted in the degradations
of various components of proteins and lipids in the gill tissue of
rainbow trout (Oncorhynchus mykiss). Exposure for 15 and 30 days
in Oreochromis niloticus (Mekkawy et al., 2013), showed that the
gill exhibited severe hyperplasia and proliferation of chloride cells
as well as the significant length shortening and reducing of the sec-
ondary gill lamellae. Subchronic exposure stages with sublethal
cadmium high-concentrations (A and B treatment) show the per-
ceived disturbance of their actin cytoskeleton. It appears in CC
located in the gill interlamellar region, at the junction between
the primary and secondary filaments. The changes correspond to
the appearance of the cytoskeleton microfilaments in the apical
part of the cell, which are attached to specific proteins in the cell
membrane. Several previous studies have shown that in various
types of mammalian cells, the process of volume control is caused
by osmotic conditions along with the emergence of drastic reorga-
nization of cytoplasmic and perinuclear actin tissue (Cornet et al.,
1993). In the shark’s rectal gland, the temporary loss of actin F-
actin is caused by hypotonicity applied to baso lateral cell surfaces
(Cooper, 1991). Since the salmon gill is involved in ion transport, it
is possible that changes in actin-induced cadmium tissue are
341
related to the actin reorganization phenomenon observed in other
tissues showing the osmoregulator properties (Izdebska et al.,
2018). Furthermore, the study of volume regulatory processes in
tumor cells indicates the role of intracellular calcium sources in
such microfilament reorganization (Cornet et al., 1993). The mem-
brane in this microfilament is what allows a small hydrophobic
motion (no affinity for the molecule) (Hertika et al., 2019).
Through passive transport (hydrophobic), the effects of cad-
mium in the cytosolic calcium concentration in fish has been
shown to be wide. In the gill epithelium (Adam et al., 2019a), cal-
cium transport is performed through CCs, as has been demon-
strated in the American eel (Flik and Verbost, 1993) and tilapia
(Flik et al., 1993). Due to its analogy to calcium, cadmium supplies
CC gills through Ca2+ pathways and after several cascade events,
they activate lipase in cytosolic Ca2+(Lin and Sung, 2003; Verbost
et al., 1994). Changes in cadmium caused by Ca2+ balance are
essential given the role played by calcium in regulating the assem-
bly and maintenance of actin structures (Janmey, 1994). This pos-
sibility is a significant cause of actin reorganization in fish treated
with pollutants (Risjani et al., 2014). In CC gills, microtubule tissue
are less developed than microfilaments. Cadmium contamination
only induces light changes in tubulin staining (Soegianto et al.,
2008). Because microfilaments and microtubule network are
highly interconnected, there is a possibility that interference with
the actin network of cadmium can affect microtubule mechanisms
as has been demonstrated in human keratinocytes in the presence
of high calcium levels (Zhang, 2005). Our results clearly show the
effects of cadmium on the network of actin microfilaments. This
raises the question of the possible impact of structural changes
on cytoskeletal function. The fractions of CC are not only involved
in calcium transport. For example, Na+/K+-ATPase lies in its broad
tubular system and is one of the critical enzymes for osmoregula-
tion. Irrespective of the possible direct effects of cadmium on the
Na+/K+-ATPase area (Izdebska et al., 2018; Schoenmakers and
Flik, 1992), the indirect impacts of metals are possible through
the relationship between enzymes and actin tissue (Lewis,
Jensen, Johnson, and Wheelock, 1995; Vandekerckhove, 1990).
Also, it is possible that actin filaments interact with other transport
systems as in different cell types (Lewis et al., 1995) or more
specifically with some temporarily activated channels such as
those occurring in the volume adjustment process (Levit, 2010).
In the presence of cadmium, the CC surface area was modified
within the first seven days, while changes in the individual density
of the CC takes longer. At the exposure stage of cadmium, CC is
derived from immature cells located in the basal epithelial layer
(Pisam et al., 1995) and is undetectable by scanning electron micro-
342 M.A. Adam et al. / Egyptian Journal of Aquatic Research 45 (2019) 337–343
scopy. The delays occurring before the density increase can be cor-
related with the rate of change of CCs and by an appearance on the
surface of the new mature cell epithelium. Furthermore, when
exposed to acidified water, marked CC proliferation has been
observed (Bonga et al., 1990).
For comparative purposes, G. affinis was exposed to cadmium to
the same degree. Experimental data suggest that action-alone
induces increased individual CC surface area, and also as a conse-
quence, such increase appear in the fractional areas. Cadmium only
causes as light increase in the cell area of the smolt cell. The max-
imum value achieved was the same as other research fish, indicat-
ing that may not exceed its responsive limits, at least at the
concentration of cadmium used in this study
Gambusia fish can adapt to osmotic pressure (Lamatsch et al.,
2015), with mechanisms required to induce various physiological
biochemical and morphological modifications (Solbakken et al.,
1996). A toxin that interferes with the transport of ions can pro-
duce hormonal effects that are reminiscent of the natural endo-
crine response. For example, cortisol and growth hormone (GH)
have been implicated in the death process (Zhang et al., 2011).
Treatment of chronic cortisol (10 days) on rainbow trout causes
in creased individual CC surface area, density and fractional area
(Prat et al., 1996). The cortisol variation is also assumed to be
related to the morphological adjustment of CC surface areas that
appear in fish subjected to various acid-base disorders (Pedersen
et al., 1999). Given the similarity between the current data and
the results obtained from acid-base disturbances or disruption pro-
cesses, it is likely that cortisol is also involved in cadmium-induced
morphological changes. Some experiments also show the GH
effects on the CC surface area, density, and extent of the opercular
and branchial fractions of CCs (Flik et al., 1993; Perry and Wood,
1985). However, this hormone is known to induce a significant
increase in the number of accessory cells. Such effects mimic what
happens naturally during the aging because of the accessory cells
associated with the mature CC (Pisam et al., 1995). The SEM study
does not indicate the emergence of accessory cells in cadmium-
administered cultures. Therefore, this is contrary to the growth
hormone implications in cadmium-induced responses. Further
experiments are needed to confirm the hypothesis implications
or normonal pathways response against cadmium pollution in
young mosquito fish and others based on differences in the level
of absorption and the ability of each organism.
Conclusion
The results of the study indicated the presence of cytotoxics in
the body of young gambusia fish exposed to cadmium. The obser-
vations made by HE staining and fluorescence illustrate the
absorption of cadmium in the fish gills of young gambusia. The
absorption affects the cytoskeleton and morphology of chloride
cells in the gills. Observations on SEM prove the change in cell
morphology. The effect of exposure to cadmium is dangerous and
toxic to the gills of young mosquito fish.
Acknowledgment
The research was funded by the Director General of Higher Edu-
cation (RISTEKDIKTI) from the Ministry of Research, Technology
and Higher Education of RI No. 158/SP2H/LT/K7/KM/2018.
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