Environmental Science and Pollution Research (2021) 28:25566–25578

https://doi.org/10.1007/s11356-021-12385-z

RESEARCH ARTICLE

The effects of some heavy metals on some fish species

Saba Naeem 1 & Muhammad Ashraf 1 & Masroor Ellahi Babar 1 & Sana Zahoor 1 & Shaukat Ali 2

Received: 8 July 2020 / Accepted: 4 January 2021 / Published online: 18 January 2021
# The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature 2021

ABSTRACT
The current research was designed to investigate the adverse effects of dense ores in different organs exposed to these metals
polluted water on three edible fish species (Wallago attu, Catla catla and Tilapia nilotica), sampled from River Sutlej at Head
Islam (Hasilpur), Pakistan. The assimilation of different elements Ni, Fe, Cd, Cr and Pb in body parts of normal fish took from the
fish farm and also made a comparison among understudied fishes. Overall absorption of ores in fishes was ranked as Fe > Ni > Cr
> Cd > Pb. A remarkable variation in the deposition of elements in different organs of three fish species and was analyzed
statistically (p < 0.05). High levels of ore assimilation in fish organs pointed out potential health risks for the fish, other aquatic
organisms and human beings too.

Keywords Fish organs . Heavy metals . River Sutlej . Histopathology

Introduction blood stream and delivered to the liver or bones as a

storage product (Obasohan 2008).
Metal concentrations have both valuable and damaging As the human population grew, the demand for the food
sound results on animals. A small number of ores are very made available to everyone has risen. The production of fish
injurious in a very little quantity while some biologically and shellfish has been much greater than before as both are the
important metals are harmful at high concentrations. first-rate supplement of protein. Internationally, humans get
Intakes of heavy metals in surplus react with bio element (25%) protein from fish and shellfish (Bahnasawy et al. 2009).
(these elements are necessary for the functioning of life) Aquatic flora and fauna have been polluted due to the trace
present in the body and destroy the structure of important element (heavy metals), the world’s major dilemma because
molecules which take part in many metabolic reactions of these elements are entering the aquatic ecosystem by various
the body (Forstner and Wittmann 1981). Heavy metal sources like the use of insecticides, pesticides, fertilizers, and
word is applied to a group of ores belonging to nonme- leaching (Fevzi 2009). World fish (75%) was used for direct
tallic that have an atomic gradient higher than five times human utilization in 2004, and it will exceed up to 98.6 % by
than water. These are called trace elements; they are pres- the year 2020 (Retnam and Zakaria 2010). The deposited el-
ent in very little quantity in the natural system. Trace ements represent absorption ratios of a known pollutant in
elements may enter the body of fish through the skin, living organisms (accumulation of specific metal in the mus-
gills, and mouth in the form of particles taken as food cle of fish) compared with that in abiotic medium (water,
by the fish. Once they are engulfed, they pass from the sediment and food). For better consideration of the deposited
factor, it is significant in forecasting the virtual assistance of a
Responsible Editor: Philippe Garrigues living medium and the deposition competence to any specific
contaminant in the viscera of fish. A high level of metal ele-
* Shaukat Ali ments in the fish body does not point to a direct toxic threat to
[email protected]
fish until there is no important accretion of metal by fish in
1 viscera (Kamaruzzaman et al. 2010).
Faculty of Science and Technology, Virtual University of Pakistan,
54-Lawrence Road, Lahore, Pakistan Heavy metals in the ecosystem are examined by detecting
2 their quantity in aqua, bottom sediments and relevant aquatic
Medical Toxicology Laboratory, Department of Zoology,
Government College University, Lahore, Pakistan life. Sediments act as vital sinks for different heavy metals and
Environ Sci Pollut Res (2021) 28:25566–25578 25567

other organic and inorganic ions. Aquatic life is at a high risk through a filter paper; finally, each sample was transferred
of absorbing pollutants which are located in the vicinity of to a plastic bottle and sealed to avoid contagion and evap-
these elements. Many species use sediments as food so the oration. All digested samples were stored at 4°C before
concentration of these poses health threats to fish, humans, heavy metal analysis. The prepared samples were ana-
and birds (Kamal et al. 2013: Edward et al. 2013). Food is lyzed for Fe, Ni, Cr, Cd, and Pb using an atomic absorp-
the main path for taking these heavy metal ions. Fish is also tion spectrometer. Metal concentration in fish tissues was
the element of this diet; it is not shocking that some of the expressed in microgram per liter of wet weight. The metal
heavy metals pass to human beings through contaminated fish analysis was performed in the Laboratory of the
(Burger and Gochfeld 2005; Shakir et al. 2013; Zeitoun 2014). Mycology Department of Botany of the University of
Punjab, Lahore.

Materials and methods

Assessment of metal accumulation in tissues
Ethical statement
At the end of the experimentation, were slaughtered for an
All trials of the animal were focused on according to the local examination of internal organs (kidneys, liver, and flesh).
and worldwide procedures. The nearby way is the Wet op de The kidneys and liver were surgically impassive and then
dierproeven (article 9) of Dutch law (international) and an assessed on a digital balance; estimation of cadmium
associated rule planned via the Bureau of Animal Research chloride concentration in the kidneys, liver, and flesh
Licensing, Local University, as detailed in our earlier papers was performed by using an atomic absorption spectropho-
(Ali et al. 2020a, b; Hussain et al. 2020; Ara et al. 2020; Khan tometer (Du Preez and Steyn 1992; Shakir et al. 2015).
et al. 2019; Ali et al. 2019; Mumtaz et al. 2019; Mughal et al. More detail is provided in our recently available paper
2019; Dar et al. 2019). (Ali et al. 2020a, b).

Area of study
Fish histopathology
River Sutlej and Head Islam Barrage: Islam Barrage is on the
River Sutlej in Hasilpur Tehsil of Bahawalpur Division of the Adult fish samples were dissected, and different tissues
Punjab province of Pakistan. It is nearly 15 kilometers (9.3 mi) (liver, gills, muscles, kidney, and heart) were removed.
north of Hasilpur and almost 7.5 kilometers south of Ludden Fats were removed and tissues were fixed. After complete
on the Vehari-Hasilpur road (Ahmadani 2017). fixation in aqueous Bouin’s solution for 24 hours, the
Fish samples were collected carefully from River Sutlej at tissues were dehydrated in an ascending series of ethanol,
Head Islam (Tehsil Hasilpur) of Division Bahawalpur of the washed with xylene, and embedded in paraffin wax.
Punjab province on the 9th of September 2018. The sampling Thick sections approximately (4–6 μm) were cut with
process was performed with the help of an expert fisherman. the help of a microtome. After de-waxing, the sections
The samples were immediately placed in a plastic case filled were stained in hematoxylin and counterstained with eo-
with river water to reduce fish mortality. Fish species were sin, dehydrated in ascending ethanol series, cleared in
packed and labeled (A, B, C), then kept in an icebox, made up xylene, and mounted in DPX (Distyrene Plasticizer
of polystyrene and brought to the laboratory within 12 h for Xylene). The slides were observed under a compound
identification following heavy metal estimation. light microscope at 100X and 400X magnification, and
Fishes were first identified from preserved specimens that photomicrographs were taken using a digital camera.
were kept in the Department of Zoology (The Islamia
University of Bahawalpur). They were labeled as fish A
(Catla catla), fish B (Wallago attu), and fish C (Tilapia Statistical data analysis
nilotica). Fish organs (liver, kidney, heart, skin epidermis,
and gills of examined fishes Catla catla, Wallago attu, and Data were presented as the mean ± standard deviation of the
Tilapia nilotica) were taken and digested by using the wet mean. Shapiro-Wilk test was used to check the distribution of
digestion method in which one gram of each wet fish tissue the data. Parametric tests were used on normally distributed
was digested with 2 ml of HNO3 and 1 ml of H2SO4 in a data and then examined statistically via one-way ANOVA,
volume metric flask. Digestion was carried out on a hot plate followed by “Dunnett’s Multiple Comparison Test”, to find
until a transparent or vine green color was attained. The any substantial variances among the group means. SPSS ver-
digested solution was diluted up to 100 ml, and from this sion 25.0 for Windows was used for analyses. Values of p ≤
50 ml of each sample was taken. Each sample was filtered 0.05 were considered significant.
25568 Environ Sci Pollut Res (2021) 28:25566–25578

Results 2013). Iron was found to be the highest in all the samples that
were analyzed. Iron was present in hemoglobin synthesis in
The concentration of heavy metals in individual red blood cells; although it was a very important element it has
organs of fish species (Catla catla, Wallago attu, and become toxic to living organisms even when exposure was
Tilapia nilotica) low. The concentration of Fe (iron) in the water sample was
also estimated that had a mean value of 0.1437 ± 0.02902 ppm
Heavy metal iron accumulation in Catla catla (Table 1 and Fig. 1). The order of occurrence of Iron in three
fishes was as a kidney of Tilapia > liver of Catla catla > gills
Iron (Fe) was present in all body parts in each fish species. of Tilapia > skin muscles of Tilapia > heart of Wallago attu
Iron deposited in the skin muscles, heart, liver, gills, and kid- (Table 1 and Fig. 1). The absorption of heavy metal Iron in the
ney of Catla catla had a mean value of 1.7877 ± 0.03485 ppm, skin muscles of three fishes was in the following order: iron
7.873 ± 0.00839 ppm, 4.0377 ± 0.20408 ppm, 1.8527 ± was significantly absorbed in the skin muscles of Tilapia
0.09624 ppm, and 0.7023 ppm, respectively (Table 1). nilotica, lower in Catla catla, and least absorbed in Wallago
There was minimum variation among the organs of the same attu.
fish. The highest concentration was recorded in the liver; it
had a mean value of 4.0377 ± 0.20408 ppm whereas the low- Bioaccumulation of heavy metal Nickel in Catla catla
est concentration was found in the heart with a mean value of
0.7873 ± 0.00839 ppm. The accumulation pattern of Fe (iron) The absorption of Ni in the body parts of Catla catla had the
in the organs of Catla catla was liver > gills > skin muscles > mean values of 0.4637 ± 0.00702 ppm, 0.3057 ± 0.00651
kidney > heart (Table 1 and Fig. 1). ppm, 0.2390 ± 0.02900 ppm, 0.2150 ± 0.00872 ppm, and
0.0937 ± 0.01464 ppm, respectively (Table 1). According to
Heavy metal iron accumulation in Wallago attu the data, the accretion order for Ni (nickel) in the organs of
Catla catla was liver > kidney > heart > skin muscles > gills.
Absorption of Fe (iron) in viscera of Wallago attu was 1.9717 It was concluded that the highest absorption has occurred in
± 0.02919 ppm, 1.8283 ± 0.02610 ppm, 1.8147 ± 0.03262 the liver and the lowest in the gills of Catla catla (Table 1
ppm, 1.678 ± 0.08372 ppm, and 1.2713 ± 0.01795 ppm, re- and Fig. 2).
spectively (Table 1). The accretion was in the rank kidney >
gills > heart > liver > skin muscles that means the kidney has
the highest absorption of iron (Fe), while the skin muscles of Bioaccumulation of heavy metal Nickel in Wallago attu
Wallago attu have the lowest absorption (Table 1 and Fig. 1).
Nickel absorption in the organs of the fish Wallago attu was
Heavy metal iron accumulation in Tilapia nilotica shown concerning (Table 1 and Fig. 2) that the heart has a
mean value of 0.3493 ± 0.02511 ppm, the liver had a mean
The kidney of Tilapia nilotica had a mean iron (Fe) concen- value of 0.2743 ± 0.01457 ppm, the kidney had a mean value
tration of 6.3207 ± 0.06676 ppm, the liver had a mean value of of 0.2233 ± 0.01258 ppm, the skin muscles had a mean value
2.4740 g ± 0.08253 ppm, the gills had a mean value of 2.355 ± of 0.1543 ± 0.01680 ppm, and the gills had a mean value of
0.12626 ppm, the muscles had a mean value of 1.8160 ± 0.1513 ± 0.02060 ppm (Table 1 and Fig. 2). The absorption of
0.01778 ppm, and the heart had a mean value of 0.0337 ± Ni (nickel) among the organs was in the following order: heart
0.00643 ppm (Table 1). So the order of (Fe) absorption was > liver > kidney > skin muscles > gills. So the heart had the
in the kidney > liver > gills > skin muscles > heart which highest absorption of nickel while the gills of Wallago attu
means that the kidney had the highest absorption of Fe had the lowest absorption (Table 1 and Fig. 2). It was evalu-
(iron) while the heart of Tilapia nilotica had the lowest ab- ated that there was minor variation in the deposition of heavy
sorption of iron (Table 1 and Fig. 1). ores among the visceral parts of three fishes under this study.

Comparative iron absorption in Catla catla, Wallago Bioaccumulation of heavy metal Nickel in Tilapia nilotica
attu and Tilapia nilotica
In Tilapia nilotica, the absorption of nickel (Ni) in gills was
Overall, the maximum absorption of Fe (iron) was found in the highest with the mean value of 0.3600 ± 0.00755 ppm,
the kidney of Tilapia nilotica as compared with Catla catla while the lowest absorption occurred in the heart as it had the
and Wallago attu (Table 1 and Fig. 1). The lowest concentra- mean value of 0.1933 ± 0.01250 ppm as mentioned (Table 1
tion was seen in the heart of Tilapia nilotica as compared with and Fig. 2); other organs had the lower amount found in the
the other two fishes. The deposition of this metal has already kidney, liver, and skin muscles, respectively, of Tilapia
been detected in the organs of freshwater fish ( Edward et al. nilotica as mentioned in Table 1. The degree of deposition
Table 1 Heavy metal absorption (ppm) in various body parts of fish species and water sample

Concentration of metals (ppm) in fish species

Catla catla Wallago attu Tilapia nilotica

Organs/ Skin Heart Liver Kidney Gills Skin Heart Liver Kidney Gills Skin Heart Liver Kidney Gills Water
Environ Sci Pollut Res (2021) 28:25566–25578

heavy muscle (M ± (M ± SD (M ± SD (M ± SD muscle (M ± (M ± SD (M ± (M ± muscle (M ± SD (M ± (M ± SD (M ± SD sample (M

metals (M ± SD SD in in ppm) in ppm) in ppm) (M ± SD SD in in ppm) SD in SD in (M ± SD in ppm) SD in in ppm) in ppm) ± SD in
in ppm) ppm) in ppm) ppm) ppm) ppm) in ppm) ppm) ppm)
Fe 1.7877 ± 0.7873 4.0377 ± 1.7023 ± 1.8527 ± 1.2713 ± 1.8147 1.6783 ± 1.9717 ± 1.8283 ± 1.8160 ± 0.0337 ± 2.4740 6.3207 ± 2.3557 ± 0.143667±
0.03485 ± 0.2040- 0.02940 0.09624 0.01795 ± 0.08372 0.0291- 0.026- 0.01778a 0.00643 ± 0.0667- 0.12626 0.029023
0.00- 8aaa 0.03- 9 10a 0.08- 6aaa
839 262 253
Ni 0.2150 ± 0.2390 0.4637 ± 0.3057 ± 0.0937 ± 0.1543 ± 0.3493 0.2743 ± 0.2233 ± 0.1513 ± 0.2457 ± 0.1933 ± 0.2687 0.3047 ± 0.3600 ± 0.031±
0.00872 ± 0.0070- 0.00651 0.01464 0.01680 ± 0.01457 0.0125- 0.020- 0.02597 0.01250 ± 0.00551 0.0075- 0.004
0.02- 2aaa 0.02- 8 60 0.01- 5aaa
900 511 343
Cr 0.0353 ± 0.0427 0.0273 ± 0.0647 ± 0.0227 ± 0.0337 ± 0.0560 0.0837 ± 0.0300 ± 0.0313 ± 0.0447 ± 0.0250 ± 0.0290 0.0147 ± 0.0347 ± 0.012667±
0.00321 ± 0.00208 0.0025- 0.00751 0.00306 ± 0.0025- 0.0010- 0.002- 0.00321 0.00265 ± 0.00351 0.00351 0.001528
0.00- 2aaa 0.00- 2aaa 0 52 0.00-
351 436 100
Cd 0.0543 ± 0.0000 0.0740 ± 0.1053 ± 0.0763 ± 0.0963 ± 0.0443 0.2560 ± 0.0673 ± 0.0240 ± 0.0517 ± 0.0780 ± 0.0710 0.0653 ± 0.1380 ± 0.064333±
0.01361 ± 0.00700 0.00950 0.00451 0.01914 ± 0.0239- 0.0106- 0.003- 0.00503 0.00900 ± 0.00208 0.0183- 0.010017
0.00- 0.00- 7aaa 0 61 0.08- 6aaa
000 321 19
Pb 0.6217 ± 0.1533 0.5797 ± 0.3933 ± 1.0113 ± 0.3723 ± 0.6680 0.5113 ± 0.3817 ± 0.6290 ± 0.3327 ± 1.0407 ± 0.3743 0.5397 ± 0.5630 ± 0.102667±
0.03320 ± 0.03814 0.06712 0.1335- 0.05052 ± 0.12799 0.0667- 0.032- 0.05100 0.1070- ± 0.11404 0.1148 0.010214
0.02- 2aaa 0.12- 1 19 6aaa 0.02-
453 900 926

“a” shows the significant difference in metal concentrations in organs of different fishes. Each value in the table represents the mean value of four duplicates and standard deviation. Arithmetical
representations: aaa = p ≤ 0.001.
25569
25570 Environ Sci Pollut Res (2021) 28:25566–25578

Fig. 1 Comparative
concentrations of iron
accumulated in fishes and river
water sample. Abbreviation and
keys: “a” shows the significant
difference in metal concentrations
in organs of different fishes. Each
bar of the graph represents the
mean value of four duplicates and
standard deviation. Arithmetical
representations: aaa = p ≤ 0.001

of Ni (nickel) was gills > kidney > liver > skin muscles > heart the liver of Catla catla and the lowest in the gills of Catla
of Tilapia nilotica (Fig. 2). catla. The order of accretion of nickel in each visceral organ of
three fishes, when a comparison among all the values was
Comparison of nickel concentration in organs of Catla made, was found in a respective order: liver of Catla catla >
catla, Wallago attu and Tilapia nilotica gills of Tilapia > heart of Wallago attu > kidney of Catla catla
> skin muscles of Tilapia nilotica (Fig. 2). The absorption of
When compared with each visceral organ of the three fishes, heavy metals in all the organs exhibited major distinction due
then the pool of Ni (nickel) was found to be the maximum in to their ability to absorb the ores. Among the three fishes,

Fig. 2 Comparative
concentrations of nickel
accumulated in fishes and river
water sample. Abbreviation and
keys: “a” shows the significant
difference in metal concentrations
in organs of different fishes. Each
bar of the graph represents the
mean value of four duplicates and
standard deviation. Arithmetical
representations: aaa = p ≤ 0.001
Environ Sci Pollut Res (2021) 28:25566–25578 25571

Catla catla showed maximum absorption. The water sample 0.00321 ppm and was low in the kidney as it had the mean
that contained these fishes was also subjected to Ni estimation value of 0.0147 ± 0.00351 ppm. There were minor changes in
that had the mean value of 0.031 ± 0.004 ppm. other organs of the Tilapia nilotica (Table 1). The concentra-
tion of chromium in the organs of Tilapia ranked as skin
Bioaccumulation of heavy metal Chromium in Catla catla muscles > gills > liver > heart > kidney (Table 1 and Fig. 3).

In Catla catla, the absorption of chromium was the highest in

the kidney with the mean value of 0.064 ± 0.00252 ppm and it Comparative absorption of chromium in Catla catla,
was the lowest in gills with the mean value of 0.0227 ± Wallago attu and Tilapia nilotica
0.00751 ppm. In other organs, there was minor variation
(Table 1). The absorption of chromium in the organs of When comparing all the mean values (Table 1), the concen-
Catla catla was in the following order: kidney > heart > skin tration of chromium in three fishes was found to be the max-
muscles > liver > gills (Table 1 and Fig. 3). imum in the liver of Wallago attu and the lowest in the kidney
of Tilapia nilotica. The level of accretion in three fish parts
was the liver of Wallago attu > kidney of Catla catla > heart
Bioaccumulation of heavy metal Chromium in Wallago attu of Wallago attu > skin muscles of Tilapia nilotica > gills of
Catla catla (Table 1 and Fig. 3). So the absorption of chromi-
Heavy metal absorption of chromium in organs of Wallago um was highly significant in the liver of Wallago attu
attu was significant in the liver as it had the mean value of (Table 1) as described earlier. Chromium absorption in water
0.0837 ± 0.00252 ppm, and chromium absorption was the was also estimated as it had a mean value of 0.0127 ± 0.00153
lowest in kidney as it had the mean value of 0.03 ± 0.001 ppm.
ppm, with little difference among other organs of the
Wallago attu (Table 1). The accretion pattern of chromium
in different body parts of Wallago attu was liver > heart > Bioaccumulation of heavy metal cadmium in Catla catla
skin muscles > gills > kidney (Table 1 and Fig. 3).
Cadmium accumulation in the organs of Catla catla was high
Bioaccumulation of heavy metal Chromium (Cr) in Tilapia in gills (Table 1) as it had a mean value of 0.0763 ± 0.00451
nilotica ppm. So it was significant. In heart of Catla catla cadmium
concentration was not significant as it had the mean value of 0
Chromium deposition in organs of Tilapia nilotica was high in ppm. Other organs of the fish have minor alterations. The
skin muscles as it had the mean value of 0.0447 ± deposition of cadmium in various parts of Catla catla was as

Fig. 3 Comparative
concentrations of chromium
accumulated in fishes and river
water sample. Abbreviation and
keys: “a” shows the significant
difference in metal concentrations
in organs of different fishes. Each
bar of the graph represents the
mean value of four duplicates and
standard deviation. Arithmetical
representations: aaa = p ≤ 0.001
25572 Environ Sci Pollut Res (2021) 28:25566–25578

follows: array gills > liver > skin muscles > kidney > heart of Tilapia > liver of Catla catla > gills of Catla catla > kidney
(Table 1 and Fig. 4). of Wallago attu (Table 1 and Fig. 4). Similar values have been
observed in the past studies too (Vinodhini & Narayanan,
Bioaccumulation of heavy metal cadmium in Wallago attu 2008). The estimation for cadmium in water was 0.0643 ±
0.01002 ppm (Table 1).
In Wallago attu, the absorption of cadmium was found signif-
icant in skin muscles with the mean value of 0.0963 ± Amassment of lead in Catla catla
0.01914 ppm and it is insignificant in gills with the mean
value of 0.0240 ± 0.00361 ppm. The kidney, heart, and liver The accumulation of lead (Pb) in the organs of fish Catla catla
have minimum variation (Table 1). The accumulation factor was significant in gills with the mean value of 1.0113 ±
was in the following sequence: skin muscles > kidney > heart 0.13352 ppm and is insignificant in the heart as it had the
> liver > gills of Wallago attu (Table 1 and Fig. 4). mean value of 0.1533 ± 0.2458 ppm (Table 1). Other organs
have little variation. The order of accumulation factor was
Bioaccumulation of cadmium in Tilapia nilotica found in the following order: gills > skin muscles > liver >
kidney > heart of fish Catla catla (Table 1 and Fig. 5).
Cadmium bioaccumulation in the organs of fish Tilapia
nilotica was estimated (Table 1). A comparatively higher con- Amassment lead (Pb) in Wallago attu
centration of cadmium was found in the heart of Tilapia with
the mean value of 0.0780 ± 0.00900 ppm, and lower absorp- In Wallago attu, the absorption of Pb (lead) was the highest in
tion was found in the gills of Tilapia with the mean value of the heart with the mean value of 0.6680 ± 0.12900 ppm. The
0.1380 ± 0.01836 ppm. Other organs (liver, kidney, and mus- lead deposition was low in the skin muscles with a mean value
cles) have minor alterations. The absorption pattern was esti- of 0.3723 ± 0.05052 ppm (Table 1). The order of lead (Pb)
mated in the following order: heart > liver > kidney > skin absorption in other parts of Wallago attu was heart > gills >
muscles > gills of Tilapia nilotica (Table 1 and Fig. 4). liver > kidney > skin muscles (Table 1 and Fig. 5).

Comparison of heavy metal cadmium in organs of Deposition of heavy metal lead (Pb) in Tilapia nilotica
Catla catla, Wallago attu and Tilapia nilotica
In Tilapia nilotica, lead (Pb) accumulation in the organs was
If compared among three fish organs, then absorption of cad- calculated (Table 1); its deposition was significant in the heart
mium was very high in skin muscles of Wallago attu > heart with the mean value of 1.0407 ± 0.10706 ppm and was the

Fig. 4 Comparative
concentrations of cadmium
accumulated in fishes and river
water sample. Abbreviation and
keys: “a” shows the significant
difference in metal concentrations
in organs of different fishes. Each
bar of the graph represents the
mean value of four duplicates and
standard deviation. Arithmetical
representations: aaa = p ≤ 0.001
Environ Sci Pollut Res (2021) 28:25566–25578 25573

Fig. 5 Comparative
concentrations of lead
accumulated in fishes and river
water sample. Abbreviation and
keys: “a” shows the significant
difference in metal concentrations
in organs of different fishes. Each
bar of the graph represents the
mean value of four duplicates and
standard deviation. Arithmetical
representations: aaa = p ≤ 0.001

lowest in skin muscles. There was a minimum alteration in Histopathology of the kidney of Catla catla
other organs of the same fish. The accumulation pattern in
different organs of Tilapia nilotica was heart > gills > kidney The kidney of Catla catla renal tubules and Bowmen’s cap-
> liver > skin muscles (Table 1 and Fig. 5). sule was observed normally. Heavy metal deposition modified
the renal tubules a condition called atrophy (a body organ or
tissue wastes away). Inflammatory cells became aggregated
Comparison of lead absorption in organs of Catla with degeneration of renal (Fig. 7).
catla, Wallago attu and Tilapia nilotica
Histopathology of the liver of Wallago attu
When compared among the three fishes, the concentration of
lead was highly significant in the heart of Tilapia nilotica with
Liver cells (Fig. 8) showed normal hepatocytes with a central-
the lowest concentration in heart of Catla catla. The sequence
ly located nucleus. While the affected cells showed cytoplas-
of deposition was the heart of Tilapia nilotica > gills of Catla
mic vacuolization, chord cells had become separated, and
catla > liver of Catla catla > skin muscles of Catla catla >
lipofuscin pigments were present between the hepatocytes
kidney of Tilapia nilotica (Table 1 and Fig. 5). Other re-
(Fig. 8).
searchers had also shown the accumulation of heavy metals
in freshwater fishes (Cirrhinus mrigala, Labeo rohita, and
Catla catla) (Shakir et al. 2015). The mean concentration of Histopathology of the liver of Tilapia nilotica
lead in water was 0.1027 ± 0.01021 ppm.
The liver of the control fish was covered with a single layer of
myoepithelial cells. The hepatic parenchyma was not arranged
Histopathology analysis into distinct lobules. The biliary channels and vascular ele-
ments exhibited the classical triads and these structures
Histopathology of the kidney of Wallago attu seemed to be randomly dispersed throughout the parenchyma,
and the hepatocytes spread out as irregular cords around the
Histology of kidney cells (control) has been shown in Fig. 6. central vein. The affected cells showed vacuoles of different
Kidney cells consisted of a glomerulus and were surrounded sizes appeared in the liver cells. Cell death, hemorrhage, and
by Bowmen’s capsule and had renal tubules. Affected kidney bursting of blood cells between the hepatocytes and dilation in
cells (Fig. 6) due to heavy metal deposition resulted in the hepatoportal blood vessels were observed. Normal fish has
expanded glomerulus and Bowmen’s capsule lost, and there hepatocytes with granular cytoplasm nucleated. Sinusoids
was a dislocation of epithelial cells. were present. Affected liver cells showed cytoplasmic
25574 Environ Sci Pollut Res (2021) 28:25566–25578

Fig. 6 Histopathology of kidney

cells of Wallago attu. a Normal
kidney cells with the glomerulus,
Bowmen’ capsule, renal tubules,
and dislocation of epithelial cells;
b abnormal kidney cells with the
expanded glomerulus. No
bowmen’s capsule

degeneration, melano-micrograph center, planktonic nuclei, vacuolization, necrosis, shrinkage, and parenchyma degener-
and mild necrosis. Lead absorption produced hepatocyte ation in fish.

Fig. 7 Histopathology of hepatocyte of Wallago attu. a Normal hepatocytes in which nucleus was centrally located; b abnormal hepatocytes with
cytoplasm (vacuolization). Blood sinusoids (separation of cord cells) and Lipofuscin pigment
Environ Sci Pollut Res (2021) 28:25566–25578 25575

Fig. 8. Histopathology of Kidney

cells of Catla catla. a, b Normal
kidney cells; c, d abnormal
kidney cells. Keys: RT, renal
tubules; BC, Bowmen’s capsule;
ART, atrophy of renal tubules;
AIC, aggregation of
inflammatory cells; DRT,
degeneration of renal tubules

Discussion metabolic processes. In this study, the estimated value of Fe

(6.3207 ± 0.06676 ppm in the kidney of Tilapia nilotica,
Metal pollution in water bionetwork had been a major prob- 4.0377 ± 0.020408 ppm in the liver of Catla catla, and
lem not only in the past, but it is still deteriorating water 1.9717 ± 0.02919 ppm in the kidney of Wallago attu) in fish
resources. Aquatic biota absorb these heavy metals, and organs far exceeded the WHO/FEPA suggested limits
through the food web, it goes fishing. It causes toxic effects 0.0300 ppm Edward et al. (2013). Although iron is an impor-
not only on aquatic biota but also on fish (Kaoud and EI- tant heavy metal, Fe (iron) has an affinity to become toxic to
Dahrshan 2010). In the present study, dense ore absorption the living creature even when exposed at little concentrations
and their effect were examined in fishes Catla catla, (Edward et al. 2013). Higher concentrations of Ni and Cr were
Wallago attu, and Tilapia nilotica. Histological effects of observed in (Table 1) the heart and vital part of Wallago attu
Ni, Fe, Cd, Cr, and Pb were observed in the internal organs while cadmium absorption (Table 1) was higher in the skin
of these fishes. Our studies showed that fishes are the most muscles of wallago attu. There was considerable variation in
sensitive to aquatic toxicants and they are the best bio absorption of dense ores in different fishes as well as in their
indicators for an aquatic environment. In the recent past, organs because this accumulation is very much dependent on
Shakir et al. (2015) have found heavy metal accumulation in their concentration in water as well as physical and chemical
five organs of Labeo rohita, Cirrhinus mrigala, and Catla qualities of water (Rauf et al. 2009).
catla captured from the different industrialized areas of In our findings, the bioaccumulation of dense ores was in the
Lahore. Similarly, Govind and Madhuri (2014) found toxic sequence Fe > Cd > Pb > Cr > Ni (Table 1). It means that
effects of dense ores in fish and higher animals. accumulation levels varied quite a lot from metal to metal, from
Current studies showed all the three fishes absorbed the fish species to species, and from organ to organ. There was a
dense ores variably in various body parts. Iron (Fe) absorption persistent increase in the absorption of heavy metals from organ
in Tilapia nilotica was in the order of Fe > Ni > Cr > Cd > Pb to organ which may be harmful to the consumers risking their
(Table 1). Our findings are in line with that of (Mahboob et al. health and well-being. The permissible limits as suggested by
2016) who found the Cu, Cr, and Fe absorption in the gills, WHO/FEPA for cadmium, iron, and lead are 0.300 ppm for Fe,
muscles, kidney, and liver of Cyprinus carpio and Wallago 0.010 ppm for Pb, and 0.003 ppm for Cd (Edward et al. 2013).
attu with comparatively higher concentrations of iron in the In our findings in the liver of catla catla, heavy metal
liver of Wallago attu. Iron (Fe) plays a major role in hemo- affected hepatocytes with cytoplasmic deterioration, scattered
globin synthesis and is a part of red blood cells. It is an im- small nuclei with mild necrosis. Our studies corroborate well
portant element of human nutrition and participates in with those of Fabio et al. (2016) who examined the
25576 Environ Sci Pollut Res (2021) 28:25566–25578

bioaccretion of cadmium, chromium, zinc, lead, and mercury Mohamed (2008) who studied bioaccumulation of selected
as well as their effects on the muscles, liver, and spleen of metals and histopathological alterations in tissues of
catfish species in Brazil. Renal tubules were atrophied and Oreochromis niloticus and Lates niloticus (Nile perch) from
deteriorated, and there was an accumulation of inflammatory Lake Nasser Egypt. Heart muscles (myocardial fibers) of
cells in the kidney. Further to this in current studies, heavy Tilapia nilotica showed hemolysis and hemorrhage with the
metal accumulation altered the morphology of the kidney of aggregation of inflammatory cells and vacuolar degeneration.
Wallago attu. The glomerulus expanded and the Bowmen Skin muscles of Tilapia nilotica degenerated and showed ne-
capsule degenerated. Hussain et al (2019) have observed sim- crosis, muscle fibers split, and muscle bundles were atrophied.
ilar histopathological changes in Wallago attu and Cirrihinus Gills of Tilapia nilotica proliferated (gill filament and second-
mrigla when they were exposed to environmentally induced ary gill lamellae) with edema in secondary gill filament. There
nephrotoxicity. Liver cells (hepatocytes) showed visible was necrosis in the epithelium of the gill filament. Mucous
vacuolization and separation of blood cord cells with the cells proliferated, secondary gill lamellae deformed, and
presence of lipofuscin pigment. Our studies are well verified curled blood vessels dilated, and there was hemorrhaging in
by that of Kaur et al. (2018) who worked on histopathology of gill filaments. Such histopathological alterations have also
Labeo rohita and Cyprinus carpio (liver, muscles, and kid- been studied by Kaoud and EI-Dahrshan (2010).
ney) on the deposition of heavy metals in their bodies.
Histopathological alterations due to heavy metals were also
described by Fatima and Usmani (2013) in the viscera of Conclusions
Channa striatus and Heteropneustes fossils.
Lead (Pb) was heavily absorbed in the skin muscles of The investigation of this research provides important informa-
Catla catla and lower in Wallago attu and the least was tion on dense ores deposition in fish taken from River Sutlej.
absorbed in Tilapia nilotica; other metals were least absorbed Fish liver, kidney (nonedible parts) and skin muscles (edible
in skin muscles of Catla catla. Javed (2005) had similar ob- part) showed higher deposition of heavy metal as a contrast to
servations on Catla catla and Labeo rohita. It had been further other body parts. Bioaccretion of dense ore in the organs is
observed in the previous studies that lead (Pb) absorption in due to bottom sediments and aquatic flora as fish feeds on
fish disturbed the immune system and fish became more vul- these elements. Heavy metal deposition decreases the homeo-
nerable to other infections (Authman et al. 2015). Similar stasis of fish. The absorption of studied dense ores in River
research had been carried out in Labeo rohita, Catla catla, Sutlej was in the sequence of Fe > Ni > Cr > Cd > Pb; these
and Cirrhinus mrigala (Javed 2005). Chromium absorption metals were deposited in different organs of fishes Catla catla,
was the highest in the kidney of Catla catla and was lower Wallago attu, and Tilapia nilotica.
in that of other fishes and followed the following pattern:
kidney of Wallago attu > kidney of Tilapia nilotica. Author Contributions Saba Naeem, Muhammad Ashraf, and Shaukat Ali
Chromium complexes induced renal failure and showed loss are responsible for the conceptualization. Saba Naeem, Muhammad
Ashraf, Masroor Ellahi Babar, Sana Zahoor, and Shaukat Ali are assigned
of osmoregulation with the induction of respiratory distress. to the data curation. Saba Naeem, Muhammad Ashraf, and Shaukat Ali
Cadmium caused epithelial swelling of renal tubules which did the formal analysis. Saba Naeem, Muhammad Ashraf, Masroor Ellahi
has also been reported by Authman et al. (2015). Babar, Sana Zahoor, and Shaukat Ali acquired the methodology.
Dense ores were not significantly absorbed in gills of Muhammad Ashraf and Shaukat Ali are responsible for the supervision.
Saba Naeem, Muhammad Ashraf, Masroor Ellahi Babar, Sana Zahoor,
Wallago attu which is in agreement with the work of and Shaukat Ali wrote the original draft. Saba Naeem, Muhammad
Mahboob et al. (2016). Heavy metal chromium deposition Ashraf, and Shaukat Ali are responsible for the review and editing.
caused vacuolization (Fatima and Usmani 2013).
Hepatocytes had become loosely arranged. Histolysis and dis- Data availability Most of the data generated during this study are includ-
integration of cell boundaries have also been observed by ed in this article. However, raw datasheets and histopathological figures
are available from the corresponding as well as co-authors upon reason-
Muthukumarvel and Rajaraman (2013). Deposition of cadmi- able request.
um affects the reproduction rate and affects the kidney which
leads to persistent toxicity (Afshan et al. 2014).
Compliance with ethical standards
Bioaccretion of dense ores in Tilapia nilotica causes sev-
eral modifications in the internal organs. The liver, heart, gills, Competing interests The authors declare no any kind of financial and
and skin muscles were badly affected by Pb, Cr, Cd, Ni, and authorship conflict of interest.
Fe. Similar to our studies, Afshan et al. (2014) also worked on
the effects of various dense ores on fish. In our study, liver Ethical approval All trials of animal were focused conferring to the local
and worldwide procedures. The nearby way is the Wet op de dierproeven
hepatocytes showed a focal area of necrosis, hemolysis of
(article 9) of Dutch law (international) and related rule scheduled via the
hepatocytes, blood vessels rupture, and thrombosis Bureau of Animal Research Licensing, Local University, as comprehen-
formation. Our investigations are also quite in line with sive in our previous articles.
Environ Sci Pollut Res (2021) 28:25566–25578 25577

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