Salient Alterations in Hepatic and Renal Histomorphology ...manuscript.advancejournals.org/uploads/3b4eca3fc52... · price in the fish market compared to some ... L. bata resolution

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.



3

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



4

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



5

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)



6

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



7

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



8

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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