Salient Alterations in Hepatic and Renal Histomorphology...
Transcript of Salient Alterations in Hepatic and Renal Histomorphology...
The International Journal of Earth & Environmental Sciences
IJEES 03|Volume 1|Issue 1|2015
1
Research Article
SALIENT ALTERATIONS IN HEPATIC AND RENAL
HISTOMORPHOLOGY OF AN INDIAN MINOR CARP, LABEO BATA
(HAMILTON, 1822) OWING TO ZNS NANOPARTICLE INDUCED
HYPOXIA AND ENVIRONMENTAL ACIDIFICATION
Nilanjana Chatterjee1, Baibaswata Bhattacharjee
2
1Department of Zoology, Ramananda College, Bishnupur-722122, Bankura, India 2Department of Physics, Ramananda College, Bishnupur-722122, Bankura, India
Correspondence should be addressed to Baibaswata Bhattacharjee
Received May 18, 2015; Accepted July 03, 2015; Published July 06, 2015;
Copyright: © 2015 Nilanjana Chatterjee et al. This is an open access article distributed under the Creative
Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Cite This Article: Chatterjee, N., Bhattacharjee, B.(2015Salient Alterations in Hepatic and Renal Histomorphology of an Indian Minor Carp, Labeo Bata (Hamilton, 1822) Owing to Zns Nanoparticle Induced Hypoxia and
Environmental Acidification. The International Journal of Earth & Environmental Sciences, 1(1).1-9
ABSTRACT
Due to enhanced surface photo-oxidation property of ZnS in its nanoparticle form, the dissolved oxygen content and pH
value of water was found to reduce in a dose dependent manner from their normal values, when ZnS nanoparticles of
different sizes are exposed to the water in various concentrations. This property was more prominent for ZnS nanoparticles
with smaller sizes. Labeo bata, exposed to ZnS nanoparticles, responded to hypoxia with varied behavioural, physiological
and cellular responses in order to maintain homeostasis and organ function in an oxygen-depleted environment. Due to the
minimization of food uptake, the hepatic cells of L. bata were found to shrink and empty spaces generated in between them
as they used storage deposit to maintain the metabolic activity of the fish. The kidneys of the exposed fishes showed
shrinkage of glomerulus and dilution of tubular lumen due to reduction in glomerular filtration rate in oxygen depleted
atmosphere. Vacuolization and hyaline degeneration of tubular epithelium were also seen in the renal histomorphology of
L. bata when the exposure time exceeded 6 days.
KEY WORDS:ZnS nanoparticles; Photo-oxidation; Hepatocytes; Renal histomorphology; Morphometry
INTRODUCTION
Snowballing of nanotechnology and mounting uses of
nanoparticles in sundry fields of sciences [1][2][3] have
increased considerably the probability that the
nanoparticles would end up in water courses either as
chemical, medical, industrial or domestic wastes. ZnS
nanoparticles (NPs) are one of such materials that can be
found in the wastes of cosmetic, pharmaceutical and rubber
industries. Apart from the various physiological disorders
due to direct uptake of nanoparticles by the aquatic animals
through different parts of their body [4][5][6][7][8][9][10],
ZnS nanoparticles are expected to exhibit some passive
effects on aquatic environment by changing important
physicochemical parameters of water due to its property of
surface photo-oxidation [11]. Due to enhanced surface
photo-oxidation property of ZnS in its nanoparticle form,
the dissolved oxygen content in water is found to reduce in
a dose dependent manner from their normal values, when
ZnS nanoparticles of different sizes are exposed to the
water in various concentrations [8][11][12]. This property
is more prominent for ZnS nanoparticles with smaller
sizes. Consequently under the exposure of ZnS NPs, the
aquatic fauna of that particular habitat are forced to live in
an oxygen depleted atmosphere [8][11][12]. When living in
a habitat with low level of dissolved oxygen, fish respond
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to hypoxia with varied behavioural, physiological, and
cellular responses in order to maintain homeostasis and
organ function in an oxygen-depleted environment
[13][14][15][16][17][18][19].
Fish is one of the major sources of edible protein in India.
Therefore, its reproduction has acquired prime importance
to the investigators working in this area. Labeo bata is a
species of freshwater Indian minor carp, found mainly in
the rivers of India, Bangladesh and Myanmar. This species
is very common, easy to cultivate and an important target
species for the small-scale fishermen. Though this fish
species has a high nutritional value in terms of protein and
micronutrients, yet it is available in a relatively cheaper
price in the fish market compared to some other fishes
having equivalent nutritional values. These reasons make
L. bata a very attractive candidate for aquaculture in the
South East Asia.
The aim of our present study is to monitor systematically
the adverse effect of ZnS NPs on histomorphology of liver
and kidney of L. bata. This will also help to realise how the
growth and maturity of the fish are being hampered when
exposed to ZnS NPs. The changing behaviour in growth
and maturity of any member of an aquatic environment due
to exposure of nanoparticles may cause an adverse effect
on the aquatic ecosystem as a whole. In the present case, it
also has its detrimental effect on the commercial market of
this fish.
EXPERIMENTAL
Synthesis and characterization of ZnS nanoparticles
(NPs)
ZnS NPs were synthesized employing simple wet chemical
method as described by Chen et al. [20]. After synthesis
the nanoparticles were characterized through Transmission
electron microscopy (TEM), Particle size analysis (PSA),
X-ray diffraction (XRD) study, Energy dispersive X-ray
(EDX) study and X-ray photo electron spectroscopy (XPS)
study. The process of synthesis and characterization
procedures of the ZnS NPs were described in detail
elsewhere [8][12]. Different characterization techniques
ascertained undoubtedly that stoichiometric, spherical ZnS
nanoparticles of different sizes (3 nm, 7 nm, 12 nm and 20
nm) were acquired under different experimental conditions
of synthesis technique[12].
Fish husbandry
Matured L. bata specimens of both sex groups caught by
means of traditional fishing gear cast net and conical trap
during daytime (10:00-15:00 hours) in monthly basis from
different places of Hooghly and Bankura districts of West
Bengal, India, were collected from the local fishermen
during the period of September, 2011 to August, 2013.
Immediately after collection, fishes were kept in watertight
containers containing tap water that has been allowed to
stand for a few days. A good supply of necessary oxygen is
provided by using a large shallow tank to ensure that a
large surface area of water is exposed to the air. Fishes are
maintained at 25°- 30°C of temperature to ensure the
natural environment. The fishes are fed on natural fish
foods. Small, regular supplies of food are provided. The
fishes are filtered out in every 10 days and are placed in
fresh water.
Histology and histometry
To study the hepatic and renal histology, liver and kidney
tissues are dissected out and cut into small pieces for
preservation in Bouin’s fixative for 18 hours. The tissues
are then dehydrated through ethanol, C2H5OH (GR, Merck
India) dried over activated molecular sieve zeolite 4A,
cleared in xylene and embedded in paraffin of melting
point 56°-58°C. Thin sections of 4 µm thicknesses are cut
using a rotary microtome machine. The sections are
stained with Delafield’s Haematoxylin and Eosin stain and
are observed under a compound light microscope of high
resolution and eventually photographed with a digital
camera attached to the microscope.
The morphometry of hepatic and renal tissues are done
using reticulo micrometer and ocular micrometer attached
to the compound light microscope. Each measurement was
made four times and their mean value was used for any
analysis.
Toxicity test
Fish specimens were exposed to six concentrations (100,
200, 250, 500, 750 and 1000 μg/L) of the ZnS
nanoparticles of different sizes (3 nm, 7 nm, 12 nm and 20
nm) for 6 days. Trials were conducted at various
concentrations to observe the impact of ZnS nanoparticles
on L. bata liver and kidney, comparing the hepatic and
renal histomorphology of the exposed fishes to that of the
fishes lived in controlled conditions. Electronic lab meters
with accuracy up to one decimal point were used to
measure the dissolved oxygen content and pH of the water.
Statistical analysis
All data were expressed as means ± SE. One-way analysis
of variance was run to compare the differences between
groups treated under different experimental conditions and
control groups. Differences were considered statistically
significant when p < 0.05. Pearson’s correlation
coefficients (r) were calculated to determine the
correlation, if any, between different hepatic and renal
morphometric parameters and nanoparticle concentrations
and exposure times at a significance level of 5%. Negative
r values prefixed by negative (-) sign and positive values
without any prefix are used in the manuscript. Curve fitting
to the experimentally obtained data was done using Origin
9.1.
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Table 1:Fitting parameters for the curves depicting the changes in the values of hepatic cell diameter (δ) with increasing
nanoparticle concentration (σ) for nanoparticles of different sizes (d) having fixed exposure time of 6 days in female L. bata
Table 2:Fitting parameters for the curves depicting the changes in the values of glomerular diameter (D) with increasing
nanoparticle concentration (σ) for nanoparticles of different sizes (d) having fixed exposure time of 6 days in female L. bata
RESULTS AND DISCUSSIONS
ZnS NP induced hypoxia and environmental
acidification
In the present study, the dissolved oxygen content in water
(DO2) was measured to be 13.2 mg/L at 15°C before any
nanoparticle was introduced in it. This value was found to
decrease both with increasing nanoparticle concentration
as well as nanoparticle exposure time in water at the same
temperature. The value of dissolved oxygen content in
water reached to as low as 3.9 mg/L for nanoparticles of
size 3 nm at a concentration of 1000 μg/L and exposure
time of 6 days.
The photo-oxidation of the surface of ZnS NPs using the
dissolved oxygen of water under sunlight and consequent
reduction of dissolved oxygen content in water has been
confirmed from detailed study of S 2p core level X ray
photoelectron spectra of ZnS nanoparticles after different
time of exposures [12]. During the surface photo-oxidation
process of ZnS NPs, The S atoms exposed to the ZnS
surface got oxidized and an increase in concentration of
chemisorbed SO2 at ZnS surface with increasing exposure
time was observed in the samples [12]. The oxide leaves
the surface as a molecular species (SO2), leaving Zn and a
freshly exposed layer of ZnS behind. Water may dissolve a
part of the SO2 released in the process causing reduction in
the pH value of the water [11]. Subsequently under the
exposure of ZnS NPs, the aquatic fauna of that particular
habitat were forced to live in an oxygen depleted and
acidified atmosphere [8][11][12].
In the present study, the pH value of water was found to
decrease when exposed to ZnS NPs in a dose dependent
manner for a fixed exposure time of 6 days. In controlled
condition the pH value of the water used in this experiment
was measured to be 7.6. This value was found to decrease
both with increasing nanoparticle concentration as well as
nanoparticle exposure time in water for a fixed
nanoparticle size. The rate of reduction in pH value was
found to be higher for the nanoparticles with smaller sizes.
In our experiment, the pH value of water dwindled down
to 4.8 for nanoparticle concentration (σ) of 1000 μg/L with
size (d) 3 nm and exposure time (t) of 6 days. Reduction of
water pH and consequent acidification of the environment
finally lead the fishes to metabolic acidosis.
After the exposure of the ZnS NPs in the water, the Zn/S
ratio in the nanoparticles was found to rise over that of the
stoichiometric value of the freshly prepared samples
confirming the loss of S from the surface of the
nanoparticles. Surfaces of the ZnS NPs, exposed to water
and light, were thus effectively destroyed by the redox
cycles and resulted in the reduction of the dissolved
oxygen content and pH value of water. This property was
found to be more prominent for ZnS NPs with smaller
sizes. This observation could be explained by the fact that
smaller particle size culminated higher surface to volume
ratio of the nanoparticles present in the water. Therefore,
ZnS NPs having smaller sizes offered greater surface area,
making the particles more sensitive to surface photo-
oxidation process. This lead to a faster deficit in dissolved
oxygen content and reduction in pH values when exposed
to water compared to the samples having larger particle
sizes.
Hepatic histology
The liver cell structure of teleosts responds very sensitively
to environmental changes, e.g. in temperature, season,
feeding conditions or presence of various chemicals in the
Nanoparticle
size (d) (nm)
δ0 (μm) α (μm) τ (μg/L) Reduced χ2
3 9.380 11.103 167.134 1.540
7 10.060 10.296 211.027 1.405
12 14.247 6.138 237.224 0.438
20 14.564 5.798 355.829 0.275
Nanoparticle size (d) (nm)
D (μm) A (μm) T (μg/L) Reduced χ2
3 49.167 48.063 500.844 0.185
7 57.468 38.327 665.272 0.492
12 59.257 36.869 911.404 0.105
20 56.990 39.477 1393.630 0.071
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water [21]. Therefore, liver histology can be used as an
indicator to show the harmful effect of ZnS NPs on L.
bata.Error! Reference source not found.(a) shows the
position of liver in a female L. bata.
Error! Reference source not found.(b) shows the
histomorphology of L. bata liver in controlled condition
portraying the liver cells in normal and healthy states. In
this figure, liver cells were found to be large with regular
outlines. These cells were dominated by storage deposits.
The nuclei were found to be large and centrally located
indicating the normal condition of the cells. The cells were
found to be in close contact, almost no empty space was
found between the cells.
Error! Reference source not found.(c)-1(e) show the effect
of increasing nanoparticle concentration on the liver
histology of L. bata. For exposure to ZnS concentration of
100µg/L (Error! Reference source not found.c), few cells
were found to be in degenerating states without a
prominent nucleus and having diffused cytoplasmic
contents. For higher concentration of ZnS nanoparticles
(σ= 500 μg/L), decrease in cell sizes due to drastic loss of
storage deposits was observed (Error! Reference source not
found.d). Therefore, the relative share of nucleus in cell
volume was strongly increased. The cells were found to be
in increasing isolated states having no close contact
between them (Error! Reference source not found.d).
Under high concentration exposure (σ= 1000 μg/L) of
smaller ZnS nanoparticles, some of the livers also showed
disruption of hepatic cell cords and apoptotic changes such
as chromatin condensation and pyknosis as indicated by
arrows in figures (Error! Reference source not found.e).
The histological alterations were more pronounced for
exposure to nanoparticles of smaller sizes. The observation
is similar for male L. bata.
Hepatic morphometry
Error! Reference source not found. shows the change in the
values hepatic cell diameter (δ) for female fishes with
increasing nanoparticle concentration (σ) for nanoparticles
of different sizes (d) used, when the exposure time is fixed
(t = 6 days). δ values are found to decrease with increase in
σ value up to 500μg/L for every size of the nanoparticles
(d) used and a fixed (t = 6 days) exposure time. Beyond
this concentration, this value remains nearly constant.
Strong negative correlation (r = -0.798) was obtained
between δ and σ for constant d and t. Analysis of
covariance revealed significant differences between the δ
values (p<0.001) for nanoparticle exposures of different
concentrations.
A significant negative correlation (r = -0.902) was revealed
between NP exposure time and hepatosite sizes during the
toxicity test. Also a significant negative correlation (r = -
0.843) could be demonstrated between exposure time and
hepatosite density for a fixed nanoparticle concentration.
The percentage of empty space in the hepatic tissue lay out
was found to increase (r = 0.712) with increasing exposure
time for a fixed concentration of ZnS NP. These
observations became more prominent with decreasing
nanoparticle sizes. Similar type of qualitative variation was
found in liver histomorphology of male L. bata.
Data presented in Error! Reference source not found.were
fitted well to the first order exponential decay curves
represented by the following equation
𝛿 = 𝛿0 + 𝛼𝑒−𝜎
𝜏 (1)
Where δ0, α and τ were the fitting parameters as shown in
Error! Reference source not found.for the family of curves
shown in Error! Reference source not found.. δ0
corresponded to the extrapolated value of hepatic cell
diameter (δ) if the nanoparticle concentration (σ) reached
infinity. The inverse of τ values determined the slopes of
the fitted curves. From the slope of the curves, it can be
established undoubtedly that the detrimental effect was
stronger for particles with smaller sizes.
These observations of alterations in hepatic
histomorphology are indicative of degradation of liver cells
under nanoparticle exposure. It has been reported that
hypoxia can induce varied behavioural, physiological, and
cellular responses among fishes
[13][14][15][16][17][18][19]. Asian dwarf striped catfish
Mystus vittatus was found to minimize their food uptake
when exposed to ZnS NP induced hypoxia [12]. Similar
pattern of altered feeding behaviour could be noticed in L.
bata in the present study. Due to the minimization of food
intake under nanoparticle exposure, the hepatic cells of the
fish were found to shrink and empty spaces generated in
between them as they used the storage in the hepatocytes
and fat vacuoles to maintain the metabolic activities of the
fishes in this adverse condition. These effects can be
associated directly with the changing feeding behaviour,
which in turn made a detrimental effect on growth,
maturity and spawning of the fish.
Renal histomorphology
The kidney is a complex organ made up of thousands of
repeating units called nephrons, each with the structure of
a bent tube. Blood pressure forces the fluid in blood to pass
a filter, called the glomerulus, situated at the top of each
nephron. In L. bata two elongated kidneys are of
mesonephric type. Error! Reference source not found.(a)
shows the position of the kidneys in female L. bata.
Error! Reference source not found.(b)-3(e) show the renal
histomorphology of L. bata under exposure of ZnS NPs of
different concentrations having size (d) of 3 nm for fixed
exposure time (t) of 6 days. The histomorphology of the
controlled kidney tissues exhibited an ordinary pattern of
renal corpuscles (consisting of glomerulus and Bowman’s
capsule) and collecting tubules with no abnormalities in
any other part of the renal cellular lay out as shown in
Error! Reference source not found.(b). When the fishes are
exposed to relatively lower concentration of ZnS NPs (σ≤
200 μg/L), the kidneys of the fishes showed shrinkage in
glomerulus and dilution of tubular lumen. For exposure to
moderate value of ZnS NPs (σ =250 μg/L), significant
decrease in glomerular size (p<0.001) and density
(p<0.001) were observed in the renal tissues of the
exposed fishes (Error! Reference source not found.c)
compared to that of the controlled fish. For exposure to
relatively higher concentration of ZnS NPs (σ = 500 μg/L),
significant decrease in the number density (p<0.001) of
collecting tubules was noticed in addition to the previous
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observations (Fig. 3d). Exposure to higher concentration of
ZnS NPs (σ≥750 μg/L), vacuolization in renal cell lay out,
hyaline degeneration of tubular epithelium was observed.
Due exposure to the highest ZnS NP concentration (σ =
1000 μg/L) used in the experiment, necrosis and dispersed
inter renal cells with pyknosis of some nuclei were
observed (Error! Reference source not found.e) in L. Bata.
Renal morphometry
Error! Reference source not found. shows the change in the
values glomerular diameter (D) for female fishes with
increasing nanoparticle concentration (σ) for nanoparticles
of different sizes (d) used, when the exposure time is fixed
(t = 6 days). D values are found to decrease gradually with
increase in σ values within the experimental limit for every
size of the nanoparticles (d) used and for a fixed exposure
time (t = 6 days). Strong negative correlation (r = -0.892)
was obtained between D and σ for constant d and t.
Analysis of covariance revealed significant differences
between the D values (p<0.001) for nanoparticle exposures
of different concentrations.
A significant negative correlation (r = -0.882) was revealed
between NP exposure time and glomerulus size during the
toxicity test, but no significant correlation could be
demonstrated between exposure time and glomerulus
density for fixed nanoparticle concentration. The lumen
diameter of the collecting tubules was found to decrease (r
= -0.704) and increase in muscular wall thickness (r =
0.801) was observed with increasing exposure time for a
fixed concentration of ZnS NP. Other time dependent
histomorphological alterations in renal tissues was not
quite prominent for relatively lower concentration of ZnS
NPs (σ<500 μg/L). When the exposure time exceeds 6
days for higher concentrations (σ≥ 500 μg/L) of ZnS NPs,
glomerular vacuolization and hyaline degeneration of
tubular epithelium were seen in the renal histomorphology
of L. bata. Similar qualitative variation was found for male
L. bata.
Data of Error! Reference source not found. are fitted well to
the first order exponential decay curves represented by the
equation
𝐷 = 𝐷0 + 𝐴𝑒−𝜎
𝑇 (2)
Where D0, A and T are the fitting parameters as shown in
Error! Reference source not found.for the family of curves
shown in Error! Reference source not found.. D0
corresponded to the extrapolated value of glomerular
diameter (D) if the nanoparticle concentration (σ) reached
infinity. The inverse of T values determined the slopes of
the fitted curves. From the slope of the curves, it can be
recognized indisputably that the harmful effect of ZnS NPs
was sturdier for particles with smaller sizes.
Ammonia is the primary metabolic waste product of most
fishes including teleosts [22][23]. Teleost freshwater fishes
occupy an environment that is hypotonic relative to their
tissues and, as a result, experience passive ion loss mainly
across the gills [24][25]. As the loss of ionic homeostasis
can lead to severe metabolic impairment [26][27][28],
teleost fishes employ mechanisms to actively take up ions,
namely Na+
and Cl-
, by reabsorbing ions across the
nephron tubules, from the glomerular filtrate back into the
blood. In addition, they actively transport ions across their
gill surfaces from the surrounding water into the blood.
Hofmann and Butler [29] reported that there exists
significant positive correlation between glomerular
filtration rate and urine flow rate in rainbow trout, Salmo
gairdneri. Glomerular filtration rate also showed linear
relationship with oxygen consumption rate of the fishes. In
the present work, ZnS NP induced hypoxia forced the
fishes to lower their oxygen consumption rate for their
metabolic activities. This is supposed to reduce the
glomerular filtration rate as well as urine flow of L. bata
under exposure to ZnS NPs. This can be attributed to the
reduction in glomerular size and density of the exposed
fishes as revealed from the histological micrographs.
Acidification of the environment due to photo oxidation of
ZnS NPs resulted in the enhancement of water H+
levels
under experimental conditions. When L. bata were
exposed to this water, the existence of H+
gradient from
water to blood generated the situation of metabolic acidosis
in the fishes reducing the blood pH level. In fish, metabolic
acidosis stimulates an elevation in ammonia excretion at
both the renal [30][31][32][33][34] and branchial
[30][31]epithelia, presumably as a means of facilitating
acid-base regulation. Reduction in water pH had been
resulted in a significant decrease in blood pH, a large
reduction in plasma HCO3 levels, a severe impairment of
swimming ability and an increase in Na+
influx in a teleost
fish Oreochromis alcalicus grahami [35].
In the present study changes in plasma acid-base status and
ionic composition along with the oxidative stress generated
by ZnS NP induced hypoxia are supposed to induce the
altered metabolic function in L. bata. This consequently
reformed the renal activity leading to the other salient
changes in renal histomorphology.
Figure 1:(a) Exposed thoracic and abdominal cavity of the female Labeo bata, showing the position of liver in situ in the thoracic region. Photomicrographs showing the liver histology of female L. bata under (b) controlled condition, (c) exposure to ZnS NP concconcentration of σ= 100 μg/L for 6 days, (d) exposure to ZnS NP concentration of σ= 500 μg/L for 6 days and (e)
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f
v
1 (a)
Li
ve
r
exposure to ZnS NP concentration of σ= 1000 μg/L for 6 days. In this case, livers tissues showed disruption of hepatic cell cords and apoptotic changes such as chromatin condensation and pyknosis as indicated by green block arrows in figure. [hepatocytes (hc), fat vacuoles (fv-white block arrows), blood vessels (Bv), empty space generated due to apoptosis ( ) and blood cells (Bc)].
Figure 2:Variation of the hepatic cell diameters (δ) against increasing nanoparticle concentrations (σ) with correspondingly fitted first order exponential decay curves for nanoparticles of different sizes (d) having fixed exposure time (t) of 6 days in female L. bata
Figure 3:(a) Exposed thoracic and abdominal cavity of the female Labeo bata, showing the position of kidney in situ in the abdominal region. Photomicrographs showing the renal histology of female L. bata under (b) controlled condition, (c) for
200 μm 200 μm
200 μm 200 μm
0 200 400 600 800 1000 12008
10
12
14
16
18
20
22
Hep
ato
cy
te d
iam
etr
e ()
(m
)
ZnS NP concentration () (g/L)
3 nm
7 nm
12 nm
20 nm
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exposure to ZnS N concentration of σ= 100 μg/L, (d) for exposure to ZnS NP concentration of σ= 500 μg/L and (e) for exposure to ZnS NP concentration of σ= 1000 μg/L *glomerulus (yellow arrow), Bowman’s capsule (Bc) and collecting tubules (ct)+.
Figure 4:Variation of the glomerular diameters (D) with increasing nanoparticle concentrations (σ) with correspondingly fitted first order exponential decay curves for nanoparticles of different sizes (d) having fixed exposure time (t) of 6 days in female L. bata.
CONCLUSION
Indian minor carp Labeo bata suffered from salient
alterations in hepatic and renal histomorphology owing to
5 mm
3 (d)
3 (b) 3 (c)
3 (e)
3 (a)
Kid
ney
5
m
m 100
μm
5 mm
0 200 400 600 800 1000 1200
50
60
70
80
90
100
3 nm
Glo
meru
lar d
iam
ete
r (
D)
(m
)
ZnS NP concentration () (g/L)
7 nm
12 nm
20 nm
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ZnS NP induced hypoxia and environmental acidification.
Due to the minimization of food intake under nanoparticle
exposure, the hepatic cells of the fish were found to reduce
in sizes generating empty spaces in between them as they
used the storage in the hepatocytes and fat vacuoles to
maintain the metabolic activities of the fishes in this hostile
condition. Onset of metabolic acidosis in the fishes as a
consequence of the environmental acidification due to the
photo oxidation of ZnS NPs resulted in the elevation in
ammonia excretion at the renal epithelia. Under the
combined effect of acidification and oxidative stress
generated by ZnS NP in the habitat, L. bata were supposed
to induce the altered metabolic function. As a result of this
reformed the renal activity, salient changes in renal
histomorphology were observed.
ACKNOWLEDGEMENT
The authors wish to thank the authority of Ramananda
College for providing some of the experimental facilities.
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