Page 1 of 26
Discoveries in Agriculture and Food Sciences - Vol. 11, No. 2
Publication Date: April 25, 2023
DOI:10.14738/dafs.112.14359.
Annamalai, A., Rajendran, N., Kaliyappan, M., & Arunachalam, M. (2023). Histopathological Changes in the Gill and Intestine Tissues
of Fish from Pichavaram Mangroves, South east Coast of India. Discoveries in Agriculture and Food Sciences, 11(2). 1-26.
Services for Science and Education – United Kingdom
Histopathological Changes in the Gill and Intestine Tissues of Fish
from Pichavaram Mangroves, South east Coast of India
Anbu Annamalai
Biodiversity and GIS Lab, Department of Environmental Sciences,
Bharathiar University, Coimbatore, Tamil Nadu, India
Narendran Rajendran
Faculty of Marine Sciences, Centre of Advanced Study in Marine Biology,
Annamalai University, Parangipettai, Chidambaram, Tamil Nadu
Manoj Kaliyappan
Tamil Nadu Forest Academy, Coimbatore, Tamil Nadu
Manimekalan Arunachalam
Biodiversity and GIS Lab, Department of Environmental Sciences,
Bharathiar University, Coimbatore, Tamil Nadu, India
ABSTRACT
A study was conducted to analysis the Histopathological Changes in the Gill and
Intestine Tissues of Mystus gulio and Mugil cephalus from Pichavaram
Mangroves for different seasons. The maximum overall metal concentrations of
Iron (Fe) recorded in the gills of M. cephalus was found to be maximum in S2
as 48.41±0.72 mg/l, copper (Cu) is maximum at station 6 as 0.52±0.02 mg/l,
chromium (Cr) accumulation in the gills of M. gulio was highest in S5 as
5.23±0.05 mg/l, the highest lead (Pb) accumulation in the gills of M. gulio was
found to be maximum in S6 as 0.71±0.03 mg/l. Likewise, the highest Iron (Fe)
accumulation in the kidney of M. gulio was found to be maximum in S2 as
36.13±0.41 mg/l followed by copper (Cu) in (S6) 0.41±0.02 mg/l, Chromium
(Cr) in (S6) records at 2.98±0.06 mg/l and by lead (Pb) maximum in S5
0.64±0.06 mg/l. The highest Iron (Fe) accumulation is found in the liver of M.
gulio in S2 as 29.14±0.34 mg/l, followed by copper (Cu) in (S3) 0.32±0.01
mg/l, Chromium (Cr) in (S6 and S1) as 2.98±0.06 & 2.98±0.03 mg/l
respectively and in lead (Pb) the highest concentration is recorded at S6
1.10±0.06 mg/l. The highest Iron (Fe) accumulation is found in the intestine
of M. gulio was found to be maximum in S6 as 29.11±0.44 mg/l and by copper
(Cu) in (S6) 1.61±0.01 mg/l and chromium (Cr) in (S5) as 2.43±0.03 mg/l and
in lead (Pb) at S6 1.74±0.03 mg/l. The highest Iron (Fe) accumulation is
found in the muscle of M. gulio in S1 as 14.03±0.37 mg/l, followed by copper
(Cu) in (S2) 1.83±0.06 mg/l, Chromium (Cr) in (S6) as 2.63±0.03 mg/l and in
lead (Pb) the maximum in S6 1.52±0.03 mg/l.
Page 2 of 26
2
Discoveries in Agriculture and Food Sciences (DAFS) Vol 11, Issue 2, April- 2023
Services for Science and Education – United Kingdom
Keywords: Pichavaram, Mystus gulio, Mugil cephalus, gills and heavy metals, Bio
accumulation.
INTRODUCTION
Industrialization has led to an exponential increase in waste generation, often discharged or
accumulated in water networks such as rivers and seas (Alesci et al., 2022). Water pollution
represents an alteration of its original characteristics through the introduction of
anthropogenic contaminants, various chemical and toxic pollutants, biocides, pesticides, and
heavy metals, in such a way as to alter its use for human food and/or for the sustenance of biotic
communities (Khan et al., 2020). The marine environment serves as a sink for contaminants
including heavy metals (Agrawal et al., 2010 & Chouvelon et al., 2012) and organic
contaminants (polychlorinated biphenyls (PCBs) and persistent pesticides) (Rajfur, 2013 &
Mormede and Davies, 2003).
India’s first and foremost occupation is agriculture and it’s an agro-based country with more
than 60–70% of its population totally reliant on agriculture. However, 30% of its agricultural
produce is lost owing to pest infestation. In the absence of a better alternative, the consumption
of pesticides becomes inevitable despite their known hazardous effects. The utilization of
pesticides in India is about 3% of the total world consumption and is increasing at the rate of
2–5% per annum (Bhadbhade et al., 2002). Heavy metals are defined as any metal or metalloid
having a relative atomic density greater than 4 g/cm3 or 5 g/cm3 that is dangerous even at very
low concentrations (Nriagu, J.O.; Pacyna, 1988 & Lenntech, 2004). Heavy metals are ubiquitous
in the environment; they are easily dissolved and carried by water, where they are quickly
absorbed by aquatic biota. Due to their high toxicity, extended persistence, and non- biodegradable nature in the food chain, heavy metals are a core group of aquatic contaminants
that cause cellular toxicity, mutagenicity, and carcinogenicity in animals; their presence in the
aquatic environment can influence water quality parameters and all forms of aquatic life
(Adeboyejo et al., 2018; Di Bella et al., 2015; Naccari et al., 2015 and Afonso et al., 2008).
Contamination of aquatic bodies with a vast array of pollutants has seriously increased
worldwide attention. Anthropogenic activities resulting from modern agricultural practices,
rapid urbanization and industrialization involve the increased release of various chemical
pollutants and toxicants, such as industrial effluents, biocides, pesticides and heavy metals etc.
which ultimately reach the aquatic environments and become responsible for their degradation
(Wang, 2002; Dautremepuits et al., 2004). Among these pollutants, heavy metals have been
recognized as strong biological poisons because of their persistent nature, toxicity, tendency to
get accumulated in organisms and undergo biomagnification (Kamble & Muley, 2000; Dinodia
et al., 2002). Heavy metals are considered the most hazardous of all environmental pollutants
(Al-Attar, 2005) due to their bioaccumulation and toxicity tendency (El-Nagger et al., 2009).
This is because heavy metals may precipitate and get absorbed on sediment particles, remain
soluble or suspended in water and/or may be taken up by aquatic fauna upon their entry into
water bodies (Mohamed, 2008). Metals are then absorbed through gills and skin and/or
ingested through food to cause bio accumulative toxicity in fish where the intensity of the
toxicity is influenced by the temperature, oxygen concentration, pH and hardness of the water
(Forstner & Wittman, 1986).
Page 3 of 26
3
Annamalai, A., Rajendran, N., Kaliyappan, M., & Arunachalam, M. (2023). Histopathological Changes in the Gill and Intestine Tissues of Fish from
Pichavaram Mangroves, South east Coast of India. Discoveries in Agriculture and Food Sciences, 11(2). 1-26.
URL: http://dx.doi.org/10.14738/dafs.112.14359
Aquatic pollution is one of the most emerging issues occurring globally and now a day water
resources have been widely exploited by the natural systems. With the rapid development of
industrialization, unplanned urbanization and an increase in human population, the pollution
of water bodies has become a universal phenomenon in the current scenario. Hence, the
protection of the coastal and marine regions from continuing pollution becomes the most
essential in coastal resources management (Lauriano et al., 2016). The evaluation of the health
status of marine fish species is a crucial step in determining an environmental assessment
(Feist et al., 2015; La Torre et al., 2020; Panebianco et al., 2021).
Fish are important organisms in the study of heavy metal pollution, because fish move freely
and assimilate heavy metals in a myriad of ways, including ingestion of suspended particles in
water, ion exchange of dissolved heavy metals through lipophilic membranes (gills), and
surface adsorption tissues and membranes. The type of exposure (dietetic or aqueous) has an
impact on the distribution of heavy metals in different fish tissues (Kaur et al., 2018).
Histopathological alterations are used as biomarkers to assess the general health of fish
exposed to pollutants (Adams, 2002). The liver, gills, and kidneys are all involved in the
accumulation and biotransformation of xenobiotics, as well as excretion and respiration in fish
(Gernhöfer et al., 2001). Because of its location, function, and blood supply, the liver is involved
in detoxification and biotransformation. It is also one of the organs most vulnerable to damage
caused by various toxic substances (Camargo & Martinez, 2007). The kidney is an important
organ for maintaining water and salt balance, for excretion of metabolic waste from the blood,
and for aspects of xenobiotic metabolism (Thophon et al., 2003 & Mabrouk, 2004). Gills are the
initial target of waterborne contaminants and are extremely sensitive to heavy metal
deposition due to constant contact with the external environment. The highly branching
morphology of gill tissues, as well as the circulation of water through them, enable heavy metal
accumulation (Shah et al., 2020). The histopathology provides a sensitive indicator of pollutant
induced stress due to the central role that the organs play in the transformation of different
active chemical compounds in the aquatic environment; particularly the gills, kidneys, and liver
are considered key organs for toxicological studies (Reish etal., 1987).
Histopathological characteristics of specific organs express condition and represent a time
integrated endogenous and exogenous impact on the organism stemming from alterations at
lower levels of biological organization (Austin, 1998). Austin (1998) reviewed the effects of
marine pollution on fish health and Au (2004) reviewed the application of histo- cytopathological biomarkers in marine pollution monitoring. Additionally, histological
biomarkers provide powerful tools to detect and characterize the biological endpoints of
toxicant and carcinogen exposure (Omar Shaik, 2012) and oil exposure (Gusma et al., 2012).
The usage of pesticides is found to be increasing use in recent years since they are
biodegradable and therefore persist in the environment only for a short time. Because of their
low persistence, repeated applications of these pesticides are being practiced for the control of
pests in agricultural fields and thereby large quantities find their way into water bodies
(Sivaperumal, 2007). A large number of pesticides are commonly used to control various
agricultural pests; however, their toxicological impact also extends to non-target species like
fish (Vinodhini and Narayanan, 2008). Fish is good indicator of aquatic contamination because
Page 4 of 26
4
Discoveries in Agriculture and Food Sciences (DAFS) Vol 11, Issue 2, April- 2023
Services for Science and Education – United Kingdom
its biochemical stress responses are quite similar to those found in mammals (Kalaiyarasi et al.,
2017). The assessment of the ecotoxicological risks caused by pesticides to ecosystems is based
on data on the toxicity and effects of pesticide preparations to non-target organisms. Fishes are
among the group of non-target aquatic organisms. Histological changes provide a rapid method
to detect the effects of irritants, especially chronic ones, in various tissues and organs (Bernet
et al., 1999). Very few histopathological studies have been reported on treatment with
dichlorvos (Bhuyan et al., 2014).
Water quality is playing an important role and it’s directly related to the safety of human health
and the animals that inhabit these environments (Moiseenko et al., 2008). Among aquatic
organisms, fishes are directly affected by an uncontrolled discharge of domestic and industrial
sewage, agricultural chemicals, heavy metals, and other xenobiotics. So, the fishes are sentinel
organisms that can be used as bioindicators of environmental stress from abiotic and biotic
changes caused by pollutants. The first responses of these organisms to environmental stress
are changes happened at the cellular and tissue levels (Svobodová, 1993). Because fish gills
have a large surface area in contact with water, structural modifications in gills have been
widely used as indicators of environmental contamination (Al-Ghanim et al., 2019).
Therefore, the present study is aimed to determine the histopathological effects on various
tissues in catfish (Mystus gulio) and mullet (Mugil cephalus) by random fishing using cast nets
in the Pichavaram mangrove forest.
MATERIALS AND METHODS
Study Area and Sampling Stations
This study was carried out in the Pichavaram mangrove forest (Lat. 11 ̊20’ N; Long. 79 ̊ 47’ E)
which is located between the Vellar and Coleroon estuaries, near Chidambaram, Tamilnadu,
India. The mangrove covers an area of about 1300 ha, of which 50% is covered by mangrove
forest, 40% by waterways and the remaining filled by sand flats and mud-flats (Krishnamurthy
and Prince Jayaseelan, 1983). The Pichavaram mangrove is influenced by the mixing of three
types of waters:
1. Neritic or costal water from the adjacent Bay of Bengal through a mouth called
‘Chinnavaikkal’,
2. Brackish water from the Vellar and Coleroon estuaries and,
3. Fresh water from an irrigation channel (Khan Sahib canal’), as well as from the main
channel of the Coleroon river.
Page 5 of 26
5
Annamalai, A., Rajendran, N., Kaliyappan, M., & Arunachalam, M. (2023). Histopathological Changes in the Gill and Intestine Tissues of Fish from
Pichavaram Mangroves, South east Coast of India. Discoveries in Agriculture and Food Sciences, 11(2). 1-26.
URL: http://dx.doi.org/10.14738/dafs.112.14359
Fig.1. Map showing the Six fishes samples collection locations in Pichavaram Mangrove
Six sampling stations were fixed along the Pichavaram Mangrove Forest. The samplinglocations
were selected based on the properties of physico chemical parameters and the source of
contaminations. The sampling locations were Backwater Zone – Mouth (N11027'13.1", E0790
47'40.1") followed by Shrimp Pond effluent site (N11025'14.4", E0790 45'56.9"), Degraded
Mangrove site (N11025'54.9", E079047'18.3"), Natural site (N11025'45.6", E079047'43.2"),
Freshwater Zone – Uppanar (N11025'41.1", E079046'15.7") and Freshwater Zone – Vellar
(N11025'04.2", E079046'19.5"). The fishes were collected from six different locations of the
Pichavaram Mangrove Forest during Post monsoon (February, 2018) and summer (May, 2018).
The fish samples were collected by using cast nets and they are collected randomly at each site.
The detail of the study area is mentioned in Figure 1. Totally, nineteen species were recorded
in Pichavaram mangrove forest in that two fish species Mystus gulio (Fig. 1a) and Mugil cephalus
(Fig. 1b) were selected based on the abundance and dominant fish catch in the four seasons.
Page 6 of 26
6
Discoveries in Agriculture and Food Sciences (DAFS) Vol 11, Issue 2, April- 2023
Services for Science and Education – United Kingdom
Fig. 2a: Mystus gulio
Fig. 2b: Mugil cephalus
The fish samples were collected and various parts like gill, intestine, liver and muscle tissue
samples were dissected from fishes immediately fixed in Bouin’s fluid for 24 h. The fixed tissues
were washed up in running tap water. Then, the tissues were dehydrated by a series of
upgraded alcohol solutions (30% alcohol for 20 min; 50% for 20 min; 70% for 30 min; 90% for
30 min; 95% for 30 min; Absolute alcohol I for 45 min and Absolute alcohol II for 45 min) and
the alcohol was cleared by a series of alcohol and xylene mixture [Alcohol (2) : Xylene (1) for
45 min; Alcohol (1) : Xylene (2) for 45 min; Xylene I for 1 h and Xylene II for 1 h] with using an
Automatic Tissue Processor and a Cold Plate (Thermo Scientific). Thin sections (5 μm sections)
were taken from the processed tissues using a microtome. The sections were floated in the
tissue flotation bath maintained at 60°C and collected on clean slides applied with Mayer’s
albumin. The sections were fixed on the slides at 60°C using a spirit lamp (Humason, 1979).
Then the slides were stained with hematoxylin and eosin stain. Histological sections are
observed under trinocular microscope. Further, the gills and intestines of the fish samples were
analyzed for histological assessments by Bernet et al. (1999).
Heavy Metal Analysis in Fish Organs
Sixty-four samples of each tissue of each fish species were collected which can accumulate
metals and yet tolerate heavy metal load. From each of the fish sample, about
15 g of gills, liver, kidney and muscles were collected and washed with distilled water,
transferred into clearly marked polyethylene flexible bags, and stored at -20oC for further
analysis. Before analysis, the tissue samples were thawed at room temperature. 1 g of sample
was weighed carefully (gills, liver, kidney and muscle) and it was digested with 5 ml Perchloric
acid and 15 ml HNO3 on a hot plate until brown fumes ceased to evolve, then samples were
cooled at room temperature, diluted with 50 ml distilled water by following (Saad et al. 2011).
Page 7 of 26
7
Annamalai, A., Rajendran, N., Kaliyappan, M., & Arunachalam, M. (2023). Histopathological Changes in the Gill and Intestine Tissues of Fish from
Pichavaram Mangroves, South east Coast of India. Discoveries in Agriculture and Food Sciences, 11(2). 1-26.
URL: http://dx.doi.org/10.14738/dafs.112.14359
Analysis of Fish Samples
Samples of gills, kidney, liver and muscle of the fishes were analyzed by following methods
described in (Saad et al. 2011) for the detection of metals viz., Pb, Cu, Fe and Cr through an
Atomic Absorption Spectrophotometer (Hitachi Polarized Zeeman AAS, Z-8200, Japan). The
blanks and calibration standard solution were also analyzed in the same way as for the samples.
The instrument calibration standards were prepared by diluting standard (1000 ppm)
purchased from Merck, Germany. A known 1000 mg/l concentration of Pb, Cu, Fe and Cr
standard solution was prepared from their salts.
Statistical Analysis
The mean values and standard error were calculated. The data were analyzed by using the
statistical package Minitab 15. Level of significance was established at P<0.05.
RESULTS
Various tests were conducted to detect the presence of heavy metals in the gills, kidney, liver
and in their intestines of Mystus gulio and M. cephalus in Pichavaram Mangrove Forest, samples
collected from six stations.
Levels of Heavy Metal Accumulation in Mystus Gulio
The maximum overall metal concentrations of Iron (Fe) recorded in the gills of M. gulio was
found to be maximum in S2 as 48.41±0.72 mg/l, followed by 26.28±0.36 mg/l (S1), 24.07±0.54
mg/l (S3), 23.34±0.33 mg/l (S4), 22.11±0.24 mg/l (S6) and minimum in 11.27±0.13 mg/l (S5)
mg/l. Likewise, the heavy metal concentration of copper (Cu) is maximum at station 6 as
0.52±0.02 mg/l (fig.4) and minimum recorded in (S1) at 0.51±0.01 mg/l. The highest chromium
(Cr) accumulation in the gills of M. gulio was found to be maximum in S5 as 5.23±0.05 mg/l,
followed by 4.24±0.06 mg/l (S6), 3.24±0.32mg/l (S4), 2.31±0.08 mg/l (S3), 1.87±0.04mg/l (S2)
and minimum in 1.24±0.05mg/l (S1) mg/l. The highest lead (Pb) accumulation in the gills of M.
gulio was found to be maximum in S6 as 0.71±0.03 mg/l, followed by 0.47±0.04 mg/l (S2),
0.39±0.06 mg/l (S3), 0.34±0.02 mg/l (S4), 0.33±0.09 mg/l (S5) and minimum in 0.20±0.02 mg/l
(S1) (Fig. 4).
Page 8 of 26
8
Discoveries in Agriculture and Food Sciences (DAFS) Vol 11, Issue 2, April- 2023
Services for Science and Education – United Kingdom
Fig. 4. Concentration of heavy metal accumulation in gills of Mystus gulio
The highest Iron (Fe) accumulation in the kidney of M. gulio was found to be maximum in S2 as
36.13±0.41 mg/l and minimum in 09.78±0.11 mg/l (S5), followed by copper (Cu) maximum in
(S6) 0.41±0.02 mg/l and minimum in 0.23±0.04 mg/l (S3). In the case of chromium (Cr), highest
in (S6) 2.98±0.06 mg/l and lowest in S1 as 1.15±0.05 mg/l and followed by lead (Pb) maximum
in S5 0.64±0.06 mg/l and minimum in 0.46±0.04 mg/l in Station 1 (Fig. 5).
Fig. 5. Concentration of heavy metal accumulation of kidney in Mystus gulio
The highest Iron (Fe) accumulation is found in the liver of M. gulio was found to be maximum
in S2 as 29.14±0.34 mg/l and minimum in 10.38±0.11 mg/l (S5), followed by copper (Cu)
maximum in (S3) 0.32±0.01 mg/l and minimum in 0.03±0.01 mg/l (S5). In the case of chromium
(Cr), highest recorded in (S6 and S1) as 2.98±0.06 & 2.98±0.03 mg/l respectively and minimum
Page 9 of 26
9
Annamalai, A., Rajendran, N., Kaliyappan, M., & Arunachalam, M. (2023). Histopathological Changes in the Gill and Intestine Tissues of Fish from
Pichavaram Mangroves, South east Coast of India. Discoveries in Agriculture and Food Sciences, 11(2). 1-26.
URL: http://dx.doi.org/10.14738/dafs.112.14359
in S4 as 2.72±0.06 mg/l and in lead (Pb) the maximum in S6 1.10±0.06 mg/l and minimum in
0.45±0.03 mg/l in Station 1 (Fig. 6).
Fig. 6. Concentration of heavy metal accumulation of liver cells in Mystus gulio
The highest Iron (Fe) accumulation is found in the intestine of M. gulio was found to be
maximum in S6 as 29.11±0.44 mg/l and minimum in 09.21±0.11 mg/l (S5), followed by copper
(Cu) maximum in (S6) 1.61±0.01 mg/l and minimum in 1.14±0.06 mg/l (S1) shown in fig.7. In
the case of chromium (Cr), highest recorded in (S5) as 2.43±0.03 mg/l and minimum in S1 as
1.08±0.06 mg/l and in lead (Pb) the maximum in S6 1.74±0.03 mg/l and minimum values were
recorded in 0.41±0.02 mg/l in Station 1&2 respectively (Fig. 7).
Fig. 7. Concentration of heavy metal accumulation in Mystus gulio intestines
The highest Iron (Fe) accumulation is found in the muscle of M. gulio was found to be maximum
in S1 as 14.03±0.37 mg/l and minimum in 12.00±0.24 mg/l (S4), followed by copper (Cu)
maximum in (S2) 1.83±0.06 mg/l and minimum in 1.32±0.03 mg/l (S4). In the case of chromium
(Cr), highest recorded in (S6) as 2.63±0.03 mg/l and minimum in S1 as 1.21±0.06 mg/l and in
Page 10 of 26
10
Discoveries in Agriculture and Food Sciences (DAFS) Vol 11, Issue 2, April- 2023
Services for Science and Education – United Kingdom
lead (Pb) the maximum in S6 1.52±0.03 mg/l and minimum values were recorded in 0.38±0.03
mg/l in Station 3 (Fig. 8).
Fig. 8. Concentration of heavy metal accumulation in the muscles of Mystus gulio
Levels of Heavy Metal Accumulation in Mugil Cephalus
The maximum overall metal concentrations of Iron (Fe) recorded in the gills of
M. cephalus was found to be maximum in S3 as 15.36±0.10 mg/l, followed by 14.48±0.09 mg/l
(S1), 14.01±0.14 mg/l (S2), 11.47±0.24 mg/l (S6), 10.24±0.24 mg/l (S4) and minimum in
8.34±0.08 mg/l (S5) mg/l. Likewise, the heavy metal concentration of copper (Cu) is maximum
in station 1 as 0.84±0.03 mg/l and minimum recorded in (S4) 0.21±0.01 mg/l. The highest
chromium (Cr) accumulation in the gills of M. cephalus was found to be maximum in S4 as
1.04±0.05 mg/l and minimum in 0.53±0.05 mg/l (S6). The highest lead (Pb) accumulation in
the gills of M. cephalus was found to be maximum in S2 & S4 as 0.38±0.04 mg/l, and minimum
in 0.18±0.02 mg/l (S1) (Fig. 9).
Page 11 of 26
11
Annamalai, A., Rajendran, N., Kaliyappan, M., & Arunachalam, M. (2023). Histopathological Changes in the Gill and Intestine Tissues of Fish from
Pichavaram Mangroves, South east Coast of India. Discoveries in Agriculture and Food Sciences, 11(2). 1-26.
URL: http://dx.doi.org/10.14738/dafs.112.14359
Fig. 9. Concentration of heavy metal accumulation in the gills of Mugil cephalus
The highest Iron (Fe) accumulation in the kidney of M. cephalus was found to be maximum in
S2 as 22.19±0.34 mg/l and minimum in 7.63±0.14 mg/l (S5), followed by copper (Cu) maximum
in (S6) 0.32±0.02 mg/l and minimum in 0.12±0.04 mg/l (S1). In the case of chromium (Cr),
highest in (S5) 0.64±0.05 mg/l and lowest in S3 as 0.24±0.03 mg/l and followed by lead (Pb)
maximum in S2 0.56±0.05 mg/l and minimum in 0.26±0.03 mg/l in Station 5 (Fig.10).
Fig. 10. Concentration of heavy metal accumulation in Mugil cephalus in kidney
The highest Iron (Fe) accumulation is found in the liver of M. cephalus was found to be
maximum in S2 as 14.36±0.41 mg/l and minimum in 7.89±0.05 mg/l (S5), followed by copper
(Cu) maximum in (S3) 0.32±0.01 mg/l and minimum in 0.09±0.01 mg/l (S5 & S6). The levels of
chromium (Cr) is highly recorded in (S3) as 0.96±0.05 mg/l and minimum in S5 as 0.35±0.03
Page 12 of 26
12
Discoveries in Agriculture and Food Sciences (DAFS) Vol 11, Issue 2, April- 2023
Services for Science and Education – United Kingdom
mg/l and in lead (Pb) the maximum in Station 60.46±0.05 mg/l and minimum in 0.15±0.03 mg/l
in Station 4 (Fig. 11).
Fig. 11. Concentration of heavy metal accumulation in liver of Mugil cephalus
The highest Iron (Fe) accumulation is found in the intestine of M. cephalus was found to be
maximum in S6 as 14.67±0.34 mg/l and minimum in 04.11±0.06 mg/l (S5), followed by copper
(Cu) maximum in (S4 & S5) 0.94±0.04 mg/l and minimum in 0.67±0.08 mg/l (S1). In the case
of chromium (Cr), highest recorded in (S5) as 0.99±0.05 mg/l and minimum in S6 as 0.37±0.02
mg/l and in lead (Pb) the maximum in S6 0.78±0.05 mg/l and minimum values were recorded
in 0.37±0.02 mg/l in Station 4 (Fig.12).
Fig. 12. Concentration of heavy metal accumulation in the intestine of Mugil cephalus
The highest Iron (Fe) accumulation is found in the muscle of M. cephalus was found to be
maximum in S1 as 14.03±0.14 mg/l and minimum in 12.01±0.05 mg/l (S6), followed by copper
Page 13 of 26
13
Annamalai, A., Rajendran, N., Kaliyappan, M., & Arunachalam, M. (2023). Histopathological Changes in the Gill and Intestine Tissues of Fish from
Pichavaram Mangroves, South east Coast of India. Discoveries in Agriculture and Food Sciences, 11(2). 1-26.
URL: http://dx.doi.org/10.14738/dafs.112.14359
(Cu) maximum in (S6) 0.98±0.05 mg/l and minimum in 0.06±0.03 mg/l (S2). In the case of
chromium (Cr), maximum recorded in (S5) as 0.87±0.05 mg/l and minimum in S1 as 0.06±0.01
mg/l and in lead (Pb) the maximum in S5 0.91±0.04 mg/l and minimum values were recorded
in 0.31±0.05 mg/l in Station 3 (Fig. 13).
Fig. 13. Concentration of heavy metal accumulation in the muscle of Mugil cephalus
Histological Analysis of Mystus gulio and Mugil cephalus
The normal fish gill of Mystus gulio was shown in fig. 14 viewed under microscope of 10x and
40x. Histology of Mystus gulio gill tissue was collected from the different locations in
Pichavaram mangrove site i.e., shrimp pond, fresh water flow areas, degrading areas of
mangroves were selected for the study showed normal histoarchitecture (Fig. 14). The gills and
intestine (Fig. 15) were consisting of Inter lamellar cell and they are composed of
undifferentiated mass of cells, justifying their high proliferative rate and supporting the
epithelial regeneration and proliferation of new mitochondria rich cells.
Fig. 14: Normal fish gill of Mystus gulio
Page 14 of 26
14
Discoveries in Agriculture and Food Sciences (DAFS) Vol 11, Issue 2, April- 2023
Services for Science and Education – United Kingdom
Summer and winter seasons showed several histological alterations like infected gills of Mystus
gulio (Fig. 15) and Mugil cephalus (Fig. 16)Infected fish intestine in Mystus gulio (Fig 18ab). The
result from the present study clearly indicates that fish gills, intestines were infected and
noticeable morphological changes were observed in their internal organs and fishes caught
nearby shrimp pond in Pichavaram Mangrove Forest are found to be slightly accumulation of
the heavy metals (18 a&b &19 a&b)
Fig. 15: Infected fish gills of Mystus gulio observed in 10 X and 40 X
Page 15 of 26
15
Annamalai, A., Rajendran, N., Kaliyappan, M., & Arunachalam, M. (2023). Histopathological Changes in the Gill and Intestine Tissues of Fish from
Pichavaram Mangroves, South east Coast of India. Discoveries in Agriculture and Food Sciences, 11(2). 1-26.
URL: http://dx.doi.org/10.14738/dafs.112.14359
Fig. 16: Infected fish gills of Mugil cephalus
Page 16 of 26
16
Discoveries in Agriculture and Food Sciences (DAFS) Vol 11, Issue 2, April- 2023
Services for Science and Education – United Kingdom
Fig. 17. Normal fish intestine observed at 10X and 40X in a. Mystus gulio & b. Mugil cephalus
Fig. 18 a&b: Infected fish intestine observed at 10X and 40X in Mystus gulio
Page 17 of 26
17
Annamalai, A., Rajendran, N., Kaliyappan, M., & Arunachalam, M. (2023). Histopathological Changes in the Gill and Intestine Tissues of Fish from
Pichavaram Mangroves, South east Coast of India. Discoveries in Agriculture and Food Sciences, 11(2). 1-26.
URL: http://dx.doi.org/10.14738/dafs.112.14359
Fig. 19 a&b: Infected fish intestine observed at 10X and 40X in Mugil cephalus
DISCUSSION
Fish are considered one of the most important biomonitors in the aquatic ecosystem for
assessing heavy metal pollution (Abou El-Gheit et al. 2012). Furthermore, fish are at the top of
the food chain and can accumulate metals that are transferred to humans, aquatic mammals
and birds through consumption of fish, causing acute or chronic diseases (Al-Yousuf et al.
2000).
Accumulation of pollutants disrupts the physiology of fish tissues. The endpoint in assessing
the risk of pollutants in the environment is the microscopic examination of target tissues
through histopathological parameters (Fatima et al. 2015). Histopathological changes can be
used as indicators of the impact of various anthropogenic pollutants on organisms and as a
measure of the overall health of the entire aquatic ecosystem (Saad et al. 2011). Harmful effects
of pollutants can be manifested in fish tissues before consequential changes in the external
appearance and behavior of fish (Mahboob et al. 2020).
In addition to different species, differences in metal concentrations depend on the types of
tissues analyzed (Abarshi et al. 2017), with gills containing a higher level of the studied metals
compared to the muscles in all tilapia species. The tissue of gills in all fish has the ability to
accumulate significant levels of metals compared to other tissues and their surface has a
negative charge and therefore provides a possible site for positively charged elements (Shovon
et al. 2017). Iron is the most important metal for biological life. It plays a greater biological role
than any other heavy metal. Its toxicity causes diarrhea, hemorrhagic gastroenteritis, liver
necrosis and leads to death by hepatic coma (Clarke et al. 1981).
Page 18 of 26
18
Discoveries in Agriculture and Food Sciences (DAFS) Vol 11, Issue 2, April- 2023
Services for Science and Education – United Kingdom
The concentration of metals, especially zinc was more elevated in the gills than in the muscles
because the gills are the main entrance for metals into the fish. They are taken up by fish directly
from water, especially through mucus and gills (Skidmore,1964). The high concentration of zinc
may be due to domestic, sewage and agricultural wastes discharged into the lake through
different drains and affecting the entire area of the lake (Abdel-Satar 2008; Abdel-Satar &
Geneid 2009; El-Naggar et al. 2016; Elmorsi et al. 2019).
Mn deficiency causes reproductive and skeletal abnormalities. Daily intake of small amounts of
Mn is recommended for growth and good health of children. However, excess consumption of
Mn can lead to neurologic and psychological disorders (Ahmed et al. 2016). Pb and Cd play no
role in biological processes of living organisms and are highly toxic non-essential elements even
at low concentrations (Dimari et al. 2008). They are also potent mutagenic and carcinogenic
agents (Markmanuel & Horsfall 2016). Pb inhibits impulse conductivity by inhibiting the
activity of acetylcholine esterase and monoamine oxidase, leading to pathological changes in
organs and tissues (Rubio et al. 1991). It also impairs the larval and embryonic growth of fish
species (Dave & Xiu, 1991).
These changes in the muscles may be attributed to the accumulation of heavy metals and/or
inorganic fertilizers that are discharged from different drains into the lake with a large amount
of wastes (Mahmoud & El-Naggar 2007; Tayel et al. 2018) and to parasitic infections (Saad et
al. 2011; Abou El-Gheit et al. 2012). The gills are the most delicate structure of the teleost body,
having an external location.
These changes may be due to fertilizers, salts and sewage discharged into Lake Al-Manzalah.
Tayel et al. (2018) and Mahmoud & Abd El Rahman (2017) found similar histopathological
changes in the liver of Mugil species and Clarias gariepinus caught in the same lake.
Degeneration and necrosis of hepatocytes may be due to the accumulation of heavy metals.
The gills and intestine of the two fish species (Mystus gulio and Mugil cephalus) collected from
mangrove areas during winter, summer, Monsoon and Post monsoon seasons the results are
exactly coincides to previous reports of Kalaiyarasi et al. (2017). Gills are the first line of defense
against waterborne toxins, and they are particularly vulnerable to heavy metal deposition due
to their constant contact with the outside world. They are also the primary site for heavy metal
uptake (Shah et al, 2020; Hermenean et al., 2017). Heavy metal intake in the gills damages the
lamella, which is involved in the ion exchange mechanism during osmoregulation (Raju, 2013).
According to Fonseca et al. (2017), metals have been linked to filament epithelium growth,
lamellar fusion, and epithelial necrosis, and their effects can be significant (Mladin et al., 2021).
Hyperplasia, lamellar fusion, epithelial necrosis, and edema have all been detected in gills and
ascribed to heavy metal toxicity (Ayoola and Alajabo, 2012).
Summer and winter seasons showed several histological alterations like infected fish gills of
Mystus gulio and infected fish gills of Mugil cephalus due to the discharge of sewage. However,
the intensity of the alterations was observed to be the highest in the specimens collected during
the summer season and the lowest during the winter season. The gills are considered to be
primary target of the contaminants as they are involved in many important functions such as
Page 19 of 26
19
Annamalai, A., Rajendran, N., Kaliyappan, M., & Arunachalam, M. (2023). Histopathological Changes in the Gill and Intestine Tissues of Fish from
Pichavaram Mangroves, South east Coast of India. Discoveries in Agriculture and Food Sciences, 11(2). 1-26.
URL: http://dx.doi.org/10.14738/dafs.112.14359
respiration, osmoregulation and excretion, remain in close contact with the external
environment, and particularly sensitive to changes in the quality of the water (8, 26). Marked
variations like hyperplasia, vacuolation, deformation of cartilage core, bubbling of gill filament,
epithelial lifting, lamellar fusion; secondary lamellar damage, shorter secondary lamellae and
erosion of secondary lamellae were noticed in the gill tissues of M. gulio and M. cephalus
collected from the polluted sites.
The gills and intestines of M. gulio and M. cephalus are drastically affected and it is clearly
noticed in histopathological studies that were observed at 10 & 40 X respectively. The gill is
playing a vital role in remodeling and it is directly related to oxygen demand and it can be
considered as a defense response that leads to a decrease in the entry of toxic compounds
through the gills (Mendes Almeida et al., 2014). Thus, it is possible that the histological changes
observed in gills follow a common induction pattern that is independent of the type of fish and
stressor agent. The increased thickness of gill filaments can act as a barrier to xenobiotics
present in water since this process could increase the distance between the capillary and the
lamellar surface, reducing the absorption of pollutants (Fig. 3b&c) (Camargo and Martinez,
2007). But, the reduced thickness of gill filaments may lead to accumulate the pollutants in
lamellar surface thereby increasing the distance between the capillary and lamellar surface.
Numerous authors have reported that increase in the numbers of these cells it may be due to
toxins exposure (Kaur et al., 2016); however, whether the origin of these histological findings
is cell division or differentiation remains unclear. The predominant pathological response of a
common fresh water fish called yellow perch (Perca flavescent) where the gills were involved
in proliferation of their basal epithelial cell and extensive proliferation in mucous cells
following changes were occurred due to increase in their salinity. This is consistent with
observations in the proliferation of gill mucous cells and basal epithelial cells following
exposure to organic contaminants (Kaur et al., 2018). Mucous cell proliferation of yellow perch
from oil sands reclaimed environments following longer residency periods has also been
observed (Nero et al., 2006), suggesting that this type of response may be a long-term
adaptation. Mucous cells contain mucins, polyanions composed of glycoproteins that can be
effective in trapping toxicants and aid in the prevention of toxicant entry into the gill epithelium
(Kaur et al., 2018). Although mucous cell proliferation may be beneficial in reducing toxicant
entry, the consequence is an increase in the distance for gas exchange along the secondary
lamellae, potentially reducing the efficiency of gas exchange and causing hypoxic conditions
(Raju et al., 2013).
When aquatic animals are exposed to toxic concentrations of heavy metals, their internal
organs may accumulate the element (Kumari et al., 2012), which may lead to biochemical and
morphological changes, particularly in the liver, intestine, gills and kidney (Abdel-Satar et al.,
2008). The present study also proves that the intestines of both the fishes were found to be
affected due to one of the reasons mentioned above. Likewise, Dar et al., 2011 reported that
catfish Clarias batrachus are exposed to 4×10-6 and 8×10-6 CdCl2 for 30 and 60 days, it includes
in the alterations in hepatocytes, eccentric positions of nuclei, enucleation, development of
cytoplasmic vacuoles, and necrosis of hepatic tissue. The uptake of metals occurs mainly
through the gills but it may also occur by means of intestinal epithelium (Mohamed, 2008).
Page 20 of 26
20
Discoveries in Agriculture and Food Sciences (DAFS) Vol 11, Issue 2, April- 2023
Services for Science and Education – United Kingdom
Toxic pollutants enter the digestive tract of fish via the food and water they consume, causing
structural and functional deterioration of the intestine (Kole et al., 2001). The present study
showed that significant damage to the intestines of both the fishes was found and shown in Fig.
4a&b. Similar findings were reported by Kaoud et al. (2011) who reported pathological changes
in the intestine of O. niloticus that were exposed cadmium, including: atrophy in the muscularis
mucosa; degenerative and necrotic changes in the mucosa and submucosa with necrotized cells
aggregated in the intestinal lumen; and edema and atrophy in the submucosa (Younis et al.,
2014). Previously by Hanna et al., 2005 reported that the changes observed in the intestine of
O. niloticus were severe degenerative and necrotic changes are noticed in intestinal mucosa.
Likewise, the present study results of the two common fish intestines (i.e., Cat fish & Mullet)
were found and the edema observed between the submucosa and mucosa may also due to the
absorption of toxic heavy metals (Hanna et al., 2005). The order of bioaccumulation of metals
in both the fishes were found to be gills was Fe> Cr > Pb > Cu; kidney and muscles were; Fe >
Cr > Cu > Pb; liver as Fe > Cu > Cr > Pb (Table 3). An increasing trend of concentration of iron,
copper, chromium and lead was observed in all the tissues. The increase of iron accumulation
is very higher in all the seasons it’s due to the discharge of wastes from the shrimp ponds and
other external factors. The level of lead, chromium and copper are in the permissible limit as
suggested by FAO (2001) are found in the tissues of fishes.
The cat fish Mystus gulio and common mullet Mugil cephalus can be suggested as a biological
model for investigating possible adaptations to pollutants. This study has confirmed the toxic
effect and the pollutants released in waters are may be due to usage of chemicals or pesticides,
etc., which are released from shrimp pond effluent sites or it may be used for agricultural
practises. Further research is in progress to check the heavy metal accumulation in the aquatic
organisms.
This is consistent with the findings of Authman & Abbas (2007) who stated that the liver is
involved in a detoxification of toxins such as heavy metals. Accumulation of hemosiderin in liver
cells may contribute to the rapid and continuous destruction of red blood cells (Hashem et al.
2020; Tayel et al. 2018; Ibrahim & Mahmoud 2005). Degeneration of hepatocytes can be caused
by oxygen deficiency due to intravascular hemolysis and vascular dilation (Gaber & Gaber
2006). Toxins secreted by microorganisms in sewage water may cause necrosis and
hemorrhage (Saad et al. 2011). Fatty degeneration can be caused by an increased rate of
utilization of energy reserves or an induced imbalance between fat utilization and production
(El-Naggar et al. 2009). The liver is an organ that excretes and binds proteins such as
metallothionein. Metal-binding proteins, which are present in the nuclei of hepatocytes,
increase the cell damage (Mela et al. 2007). The present study evidence that the sample
collected from station 6 (nearby aquaculture pond) shown high amount of heavy metal
accumulation in the internal organs of the two fish species from Pichavaram waters. These
fishes are abundantly available in all the seasons and are edible, consumed by various living
organisms including human beings and having a greater chance of accumulation of heavy metal
transfer from one trophic level to another trophic level; shortly called as Biological
Magnification. Further studies have been planned to study the toxicity of these heavy metals in
human beings residing in and around the Pichavaram.
Page 21 of 26
21
Annamalai, A., Rajendran, N., Kaliyappan, M., & Arunachalam, M. (2023). Histopathological Changes in the Gill and Intestine Tissues of Fish from
Pichavaram Mangroves, South east Coast of India. Discoveries in Agriculture and Food Sciences, 11(2). 1-26.
URL: http://dx.doi.org/10.14738/dafs.112.14359
Acknowledgements
We are very thankful to DST-SERB for the financial support (No. SB/EMEQ-060/2013; dt.
12.07.2013), Principal Chief Conservator of Forest & District Forest Officer Cuddalore Division,
Tamil Nadu Forest Department for the permission to collect the samples and Mr. R. Narendran
& Mr. P. Dinesh, Ph. D Research Scholar, Centre for Advance Studies in Marine Biology,
Annamalai University for assisting in sample collection, and for the authorities of Bharathiar
University for providing necessary laboratory facilities.
References
Abarshi, M.M., Dantala, E.O. & Mada, S.B. (2017). Bioaccumulation of heavy metals in some tissues of croaker fish
from oil spilled rivers of Niger Delta region, Nigeria. Asian Pac. J. Trop. Biomed. 7(6): 563–568.
Abdel-Satar, A.M. (2008). Chemistry of major ions, nutrient salts and heavy metals in the Manzalah Lake system,
Egypt. Egypt. J. Aqua. Res. 34(2): 130–148.
Abdel-Satar, A.M., Geneid, Y.A. (2009). Evaluation of heavy metal status in ecosystem of Lake Manzalah, Egypt.
Global. J. Environ. Res. 3(3): 194–204.
Abou El-Gheit, E.N., Abdo, M.H. & Mohmoud, S.A. (2012). Impact of blooming phenomenon on water quality and
fishes in Qarun Lake, Egypt. International Journal of Environmental Science and Engineering (IJESE) 3: 11–23.
Adams, S.M. Biological Indicators of Aquatic Ecosystem Stress; American Fisheries Society: New York, NY, USA,
2002.
Adeboyejo, A.O.; Clarke, E.O.; Hammed, A.M.; Adaramoye, R.O. Haematological and Hepatic Responses of the
African Catfish Clarias gariepinus to Sublethal Exposure of Industrial Effluents from Ologe Lagoon Environs,
Lagos, Nigeria. J. Food Sci. Eng. 2018, 8, 198–209.
Afonso, C.; Lourenço, H.M.; Pereira, C.; Martins, M.F.; Carvalho, M.L.; Castro, M.; Nunes, M.L. Total and organic
mercury, selenium and α-tocopherol in some deep-water fish species. J. Sci. Food Agric. 2008, 88, 2543–2550.
Agrawal, A.; Catalini, C.; Goldfarb, A.; NET Institute Working paper, no. The Geography of Crowdfunding; NET
Institute Working paper no. 10-08; National Bureau of Economic Research: Cambridge, MA, USA, 2010.
Ahmed, Md.K., Abdul Baki, M., Kundu, G.K., Islam, Md.S., Islam, Md.M. et al. (2016). Human health risks from
heavy metals in fish of Buriganga River, Bangladesh. Springer Plus 5: 1697
Al-Attar AM. Changes in hematological parameters of the fish, Oreochromis niloticus treated with sublethal
concentrations of cadmium. Pakistan J Biol Sci. 2005;8(3):421–4.
Alesci, A.; Cicero, N.; Fumia, A.; Petrarca, C.; Mangifesta, R.; Nava, V.; Lo Cascio, P.; Gangemi, S.; Di Gioacchino, M.;
Lauriano, E.R. Histological and Chemical Analysis of Heavy Metals in Kidney and Gills of Boops boops:
Melanomacrophages Centers and Rodlet Cells as Environmental Biomarkers. Toxics 2022, 10, 218.
https://doi.org/ 10.3390/toxics10050218
Al-Ghanim, K.A., Ahmed, Z., Al. Balwi, H.A., Al-Misned, F., Mahboob, S., Suliman, El., 2019. Accumulation and
histological transformation in the gills, liver, muscles, and skin in Oreochromis niloticus induced by mercury.
Turk. J. Vet. Anim. Sci. https://doi.org/10.3906/vet-1710-50
Alinnor IJ, Obiji IA. Assessment of trace metal composition in fish samples from Nworie River. Pakistan J Nutri.
2010;9(1):81–5.
Page 22 of 26
22
Discoveries in Agriculture and Food Sciences (DAFS) Vol 11, Issue 2, April- 2023
Services for Science and Education – United Kingdom
Al-Yousuf, M.H., El-Shahawi, M.S. & Al-Ghais, S.M. (2000). Trace metals in liver, skin and muscle of Lethrimus
lentjan fish species in relation to body length and sex. The Science of the Total Environment 256(2–3): 87–94.
Au DW. The application of histo-cytopathological biomarkers in marine pollution monitoring: a review. Mar
Pollut Bull. 2004 May;48(9-10):817-34. doi: 10.1016/j.marpolbul.2004.02.032. PMID: 15111029.
Austin B. The effects of pollution on fish health. Journal of Applied Microbiology. 1998; 85(1):234S-242S.
Authman, M. & Abbas, H. (2007). Accumulation and distribution of copper and zinc in both water and some vital
tissues of two fish species (Tilapia zillii and Mugil cephalus) of Lake Qarun, Fayoum Province, Egypt. Pak. J. Biol.
Sci. 10(13): 2106–2122.
Ayoola, S.O.; Alajabo, O.T. Acute Toxicity and Histopathological Effects of Engine Oil on Sarotherodon
melanotheron (Black Jaw Tilapia). Am.-Eurasian J. Toxicol. Sci. 2012, 4, 48–55.
Bernet, D., Schmdit, H., Meier, W., Burkhardt-Holm, P. and Wahli, T., 1999. Histopathology in fish: proposal for a
protocol to assess aquatic pollution. J. Fish Dis., 22: 25- 34.
Bhadbhade, B.J. & Sarnaik, S.S. & Kanekar, Pradnya. (2002). Biomineralization of an organophosphorus pesticide,
Monocrotophos, by soil bacteria. Journal of applied microbiology. 93. 224-34. 10.1046/j.1365-2672.2002.01680.
x.
Bhuyan, G., Anandhan, R., Kavitha, V., 2014. Qualitative and quantitative analysis of fish tissue of Oreochromis
mossambicus collected from Kedilam river Cuddalore Tamil Nadu, India. Int. J. Appl. Sci. Biotech. 2 (2), 135–141.
Camargo, M.M.P.; Martinez, C.B.R. Histopathology of gills, kidney and liver of a Neotropical fish caged in an urban
stream. Neotrop. Ichthyol. 2007, 5, 327–336.
Chouvelon, T.; Spitz, J.; Caurant, F.; Mèndez-Fernandez, P.; Autier, J.; Lassus-Débat, A.; Chappuis, A.; Bustamante,
P. Enhanced bioaccumulation of mercury in deep-sea fauna from the Bay of Biscay (North-East Atlantic) in
relation to trophic positions identified by analysis of carbon and nitrogen stable isotopes. Deep Sea Res. Part I
Oceanogr. Res. Pap. 2012, 65, 113–124.
Clarke, M.L., Harvey, D.G. & Humphreys, D.J. (1981). Veterinary Toxicology. London: Bailliere Tindall
Dautremepuits C, Paris-Palacios S, Betoulle S, Vernet G. Modulation in hepatic and head kidney parameters of
carp (Cyprinus carpio) induced by copper and chitosan. Toxicology and Pharmacology. 2004; 137:325-333.
Dave, G & Xiu, R (1991). Toxicity of mercury, copper, nickel, lead and cobalt to embryos and larval of Zebra fish
Brachydanio rerio. Archive of Environmental Contamination and Toxicology 21: 126–134.
Di Bella, G.; Potortì, A.G.; Turco, V.L.; Bua, D.; Licata, P.; Cicero, N.; Dugo, G. Trace elements in Thunnus thynnus
from Mediterranean Sea and benefit–risk assessment for consumers. Food Addit. Contam. Part B 2015, 8, 175–
181.
Dimari, G.A., Abdulrahman, J.C. & Garba, S.T. (2008). Metals concentrations in tissues of Tilapia gallili, Clarias
lazera and Osteo glossidae caught from Alau Dam, Maiduguri, Borno State, Nigeria. American Journal of
Environmental Sciences 4(4): 373–379
Dinodia GS, Gupta RK, Jain KL. Effect of cadmium toxicity on liver glycogen in some freshwater fishes. Proceeding
of XI National Symposium on Environment. 2002; 11:236-238
Elmorsi, R.R., Abou-El-Sherbini, K.S., Mostafa, G.A. & Hamed, M.A. (2019) Distribution of essential heavy metals in
the aquatic ecosystem of Lake Manzala, Egypt. Heliyon 5(8): e02276.
Page 23 of 26
23
Annamalai, A., Rajendran, N., Kaliyappan, M., & Arunachalam, M. (2023). Histopathological Changes in the Gill and Intestine Tissues of Fish from
Pichavaram Mangroves, South east Coast of India. Discoveries in Agriculture and Food Sciences, 11(2). 1-26.
URL: http://dx.doi.org/10.14738/dafs.112.14359
El-Naggar, N., Rifaat, A.E. & Khalil, M.Kh. (2016) Numerical modeling on water flow in Manzala Lake, Nile Delta,
Northern Egypt. International Journal of Contemporary Applied Sciences 3(4): 28–44.
El-Nagger AM, Mohamoud SA, Tayel SI. Bioaccumulation of some heavy metals and histopathological alterations
in liver of Oreochromis niloticus in relation to water quality at different localities along the River Nile, Egypt.
World J Fish Mar Sci. 2009;1(2):105–14.
Feist, S.; Stentiford, G.; Kent, M.; Santos, A.R.; Lorance, P. Histopathological assessment of liver and gonad
pathology in continental slope fish from the northeast Atlantic Ocean. Mar. Environ. Res. 2015, 106, 42–50.
Fonseca, A.; Fernandes, L.F.S.; Fontaínhas-Fernandes, A.; Monteiro, S.M.; Pacheco, F. The impact of freshwater
metal concentrations on the severity of histopathological changes in fish gills: A statistical perspective. Sci. Total
Environ. 2017, 599–600, 217–226.
Fonseca, A.; Fernandes, L.F.S.; Fontaínhas-Fernandes, A.; Monteiro, S.M.; Pacheco, F. The impact of freshwater
metal concentrations on the severity of histopathological changes in fish gills: A statistical perspective. Sci. Total
Environ. 2017, 599–600, 217–226.
Forstner V, Wittman GTW. Metal pollution in the aquatic environment. New York: Springer-Verlag, Berlin
Heidelberg; 1986.
Gaber, H.S. & Gaber, S.A. (2006). The effect of water quality of Lake Quarun on liver and gonads of Oreochromis
aureus (L.) and Tilapia zillii (Gev.). Egypt. J. Aquat. Res. 32(2): 335–349.
Gernhöfer, M.; Pawert, M.; Schramm, M.; Müller, E.; Triebskorn, R. Ultrastructural biomarkers as tools to
characterize the health status of fish in contaminated streams. J. Aquat. Ecosyst. Stress Recover. 2001, 8, 241–
260.
Gusma EP, Rodrigues V, Moreira CB, Romano LA, Sampaio LA, Miranda-Filho KC. Growth and histopathological
effects of chronic exposition of marine Pejerrey Odontesthes argentinensis larvae to petroleum water soluble
fraction. Royal Swedish Academy of Science. 2012; 41:456-466.
Hashem, M.H., Tayel, S.I., Sabra, E.A., Yacoub, A.M. & Heiba, A.A. (2020). Impact of the water quality of El-Rahawy.
Drain on some genetic and histopathological aspects of Oreochromis niloticus. Egypt J. Aquat. Biol. & Fish. 24(2):
19–38.
Hermenean, A.; Gheorghiu, G.; Stan, M.S.; Herman, H.; Onita, B.; Ardelean, D.P.; Ardelean, A.; Braun, M.; Zsuga, M.;
Kéki, S.; et al. Biochemical, Histopathological and Molecular Responses in Gills of Leuciscus cephalus Exposed to
Metals. Arch. Environ. Contam. Toxicol. 2017, 73, 607–618.
Humason GL. 1979. Animal Tissue Techniques, 4th ed. Freeman, San Francisco, CA, USA.
Ibrahim, S.A. & Mahmoud, S.A. (2005). Effect of heavy metals accumulation on enzyme activity and histology in
liver of some Nile fish in Egypt. Egypt. J. Aquat. Biol. & Fish. 9(1): 203– 219
Kalaiyarasi T, Jayakumar N, Jawahar P, Ahilan B and Subburaj A (2017). Histological changes in the gill and liver
of marine spotted catfish, Arius maculatus from sewage disposal site, Therespuram off Thoothukudi, Southeast
coast of India. Journal of Entomology and Zoology Studies 2017; 5(5): 1710-1715
Kamble GB, Muley DV. Effect of acute exposure of endosulfan and chlorpyriphos on the biochemical composition
of the freshwater fish, Sarothero mossambicus. Indian Journal of Environmental Sciences. 2000; 4(1):97-102
Kaur, S.; Khera, K.S.; Kondal, J.K. Heavy Metal Induced Histopathological Alterations in Liver, Muscle and Kidney
of Freshwater Cyprinid, Labeo Rohita (Hamilton). J. Entomol. Zool. Stud. 2018, 6, 2137–2144.
Page 24 of 26
24
Discoveries in Agriculture and Food Sciences (DAFS) Vol 11, Issue 2, April- 2023
Services for Science and Education – United Kingdom
Khan, M.S.; Javed, M.; Rehman, T.; Urooj, M.; Ahmad, I. Heavy metal pollution and risk assessment by the battery
of toxicity tests. Sci. Rep. 2020, 10, 16593.
La Torre, G.L.; Cicero, N.; Bartolomeo, G.; Rando, R.; Vadalà, R.; Santini, A.; Durazzo, A.; Lucarini, M.; Dugo, G.;
Salvo, A. Assessment and Monitoring of Fish Quality from a Coastal Ecosystem under High Anthropic Pressure: A
Case Study in Southern Italy. Int. J. Environ. Res. Public Health 2020, 17, 3285.
Lauriano, E.R.; Zuwała, K.; Kuciel, M.; Budzik, K.A.; Capillo, G.; Alesci, A.; Pergolizzi, S.; Dugo, G.; Zaccone, G.
Confocal ̇ immunohistochemistry of the dermal glands and evolutionary considerations in the caecilian,
Typhlonectes natans (Amphibia: Gymnophiona). Acta Zool. 2016, 97, 154–164
Lenntech, K. Water Treatment and Air Purification; Lenntech: Rotterdam, The Netherlands, 2004.
Mabrouk, D.B.M. Aspects of Manzalah Lake Pollution on Mugil Species. Ph.D. Thesis, Ain Shams University, El
Cairo, Egypt, 2004.
Mahboob, S., Al-Ghanim, K.A., Al-Balawi, H.F., Al-Misned, F. & Ahmed, Z. (2020). Toxicological effects of heavy
metals on histological alterations in various organs in Nile tilapia (Oreochromis niloticus) from freshwater
reservoir. Journal of King Saud University- Science 32(1): 970–973.
Mahmoud, S.A. & El-Naggar, A.M. (2017). Alterations in Clarias gariepinus caused by pollutants at El-Rahawy
area, Rosetta branch, River Nile, Egypt. J. Egypt. Acad. Environ. Develop.8(2): 61–70.
Markmanuel, D.P. & Horsfall, Jnr M. (2016). Evaluation of carcinogenic and non-carcinogenic risk of cadmium
and nickel in land snails (A. achatina and L. flammea) and marine snails (P. aurita and T. fuscatus) commonly
consumed in Nigeria. Acta Chim. Pharm. Indica 6(4): 123–134.
Mela, M.R., Ventura, F., Carvalho, D.F., Pelletier, C.E. & Ribeiro, C.A. (2007). Effects of dietary methylmercury on
liver and kidney histology in the neotropical fish Hoplias malabaricus. Ecotoxicology and Environmental Safety
68(3): 426–435.
Mladin, B.O.; Albu, P.; Herman, H.; Balta, C.; Lazar, V.; Fulop, A.; Baranyai, E.; Harangi, S.; Keki, S.; Nagy, L.; et al.
Correlation between Heavy Metal-Induced Histopathological Changes and Trophic Interactions between
Different Fish Species. Appl. Sci. 2021, 11, 3760.
Mohamed FAS. Bioaccumulation of selected metals and histopathological alterations in tissues of Oreochromis
niloticus and Lates niloticus from Lake Nasser, Egypt. Global Vet. 2008;2(4):205–18.
Moiseenko TI, Gashkina NA, Sharova YN, Kudryavtseva LP (2008) Ecotoxicological assessment of water quality
and ecosystem health: A case study of the Volga River. Ecotoxicol. Environ Saf 71: 837-850.
Mormede, S.; Davies, I.M. Horizontal and vertical distribution of organic contaminants in deep-sea fish species.
Chemosphere 2003, 50, 563–574.
Naccari, C.; Cicero, N.; Ferrantelli, V.; Giangrosso, G.; Vella, A.; Macaluso, A.; Naccari, F.; Dugo, G. Toxic Metals in
Pelagic, Benthic and Demersal Fish Species from Mediterranean FAO Zone 37. Bull. Environ. Contam. Toxicol.
2015, 95, 567–573.
Nriagu, J.O.; Pacyna, J.M. Quantitative assessment of worldwide contamination of air, water and soils by trace
metals. Nature 1988, 333, 134–139.
Omar Shaik AM. Histopathological Response of the Intestinal Tissue of Lignose Emperor Lethrinus elongates To
Crude Oil Exposure, Global. Advanced Research Journal of Environmental Science and Toxicology. 2012;
1(9):211-214.
Page 25 of 26
25
Annamalai, A., Rajendran, N., Kaliyappan, M., & Arunachalam, M. (2023). Histopathological Changes in the Gill and Intestine Tissues of Fish from
Pichavaram Mangroves, South east Coast of India. Discoveries in Agriculture and Food Sciences, 11(2). 1-26.
URL: http://dx.doi.org/10.14738/dafs.112.14359
Panebianco, F.; Nava, V.; Giarratana, F.; Gervasi, T.; Cicero, N. Assessment of heavy- and semi-metals
contamination in edible seaweed and dried fish sold in ethnic food stores on the Italian market. J. Food Compos.
Anal. 2021, 104, 104150.
Puttaiah ET, Kiran BR. Heavy metal transport in a sewage fed Lake of Karnataka, India. In: Edited by Sengupta M,
Dalwani R. Proceedings of Taal2007. The 12th World Lake Conference; Jaipur, India 2008.
Rajfur, M. Algae—Heavy Metals Biosorbent/Glony—Biosorbent Metali Ci ̨ezkich. ̇ Ecol. Chem. Eng. S 2013, 20,
23–40.
Raju, K.V.; Somashekar, R.K.; Prakash, K.L. Metal Concentration in Fresh Water Fish Organs. Open J. Met. 2013, 3,
23–28.
Reish, D.L.; Oshida, P.S. (Eds.) Manual of Methods in Aquatic Environment Research. 10: Short-Term Static
Bioassays/by Donald L. Reish and Philip S. Oshida; FAO Fisheries Technical Paper: Rome, Italy, 1987; ISBN 978-
92-5-102517-8.
Rubio, R., Tineo, P., Torreblance, A., Del-Romo, J. & Mayans, J.D. (1991). Histological and electron microscopical
observations on the effects of lead on gills and midget gland of Procamarus clarkii. Toxicology and Environmental
Chemistry 31(1): 347–352.
Saad, S.M.M., El-Deeb, A.E., Tayel, S.I. & Ahmed, N.A.M. (2011). Haematological and histopathological studies on
Clarias gariepinus in relation to water quality along Rossetta Branch, River Nile, Egypt. Egypt. J. Exp. Bio. (Zool.)
7(2): 223–233
Shah, N.; Khan, A.; Ali, R.; Marimuthu, K.; Uddin, M.N.; Rizwan, M.; Rahman, K.U.; Alam, M.; Adnan, M.;
Muhammad; et al. Monitoring Bioaccumulation (in Gills and Muscle Tissues), Hematology, and Genotoxic
Alteration in Ctenopharyngodon idella Exposed to Selected Heavy Metals. BioMed Res. Int. 2020, 2020, 6185231
Shovon, M.N.H., Majumdar, B.C. & Rahman, Z. (2017). Heavy Metals (Lead, Cadmium and Nickel) Concentration in
Different Organs of Three Commonly Consumed Fishes in Bangladesh. Fish. Aqua. J. 8: 207. DOI: 10.4172/2150-
3508.1000207.
Sivaperumal P, Sankar TV, Viswanathan Nair PG (2007) Heavy metal concentrations in fish, shellfish and fish
products from internal markets of India vis-a-vis international standards. Food Chem 102: 612-620.
Skidmore, J. (1964). Toxicity of zinc compounds to aquatic animals with special references to fish. Quart. Rev.
Biol. 39(3): 227–248.
Svobodová Z (1993) Water Quality and Fish Health. FAO, Rome, EIFAC technical paper No. 54, 67pp.
Tayel, S.I., Mahmoud, S.A., Ahmed, N.A.M. & Abdel Rahman, A.S. (2018). Pathological impacts of environmental
toxins on Oreochromis niloticus fish inhabiting the water of Damietta branch of the River Nile, Egypt. J. Aquat.
Biol. & Fish. 22(5): 309–321
Thophon, S.; Kruatrachue, M.; Upatham, E.; Pokethitiyook, P.; Sahaphong, S.; Jaritkhuan, S. Histopathological
alterations of white seabass, Lates calcarifer, in acute and subchronic cadmium exposure. Environ. Pollut. 2003,
121, 307–320.
Vander Oost R, Beyer J, Vermeykebm NPE. Fish bioaccumulation and biomarkers in environmental risk
assessment: a review. Environ Toxicol Pharmacol. 2003; 13:57–149.
Vinodhini R, Narayanan M (2008) Bioaccumulations of heavy metals in organs of fresh water fish Cyprinus carpio
(Common carp). Int J Environ Sci Tech 5: 179-182.
Page 26 of 26
26
Discoveries in Agriculture and Food Sciences (DAFS) Vol 11, Issue 2, April- 2023
Services for Science and Education – United Kingdom
Wang WX. Interaction of trace metals and different marine food chains. Marine Ecology Progress Series. 2002;
243: 295-309.
Mendes Almeida, Patríca Natalícia; Silva Freitas, Cláudio Eduardo; Oliveira Abrão, Flávia; Oliveira Ribeiro,
Izabella Carolina; Alves Vieira, Edvaldo; Castro Geraseev, Luciana; Robson Duarte, Eduardo Atividade.
Cellulolytic activity of aerobic fungi isolated from dairy cattle fed with forage tropical. Revista Caatinga, vol. 27,
núm. 4, octubre-diciembre, 2014, pp. 202-207
Kaur, S.; Khera, K.S.; Kondal, J.K. Heavy Metal Induced Histopathological Alterations in Liver, Muscle and Kidney
of Freshwater Cyprinid, Labeo Rohita (Hamilton). J. Entomol. Zool. Stud. 2018, 6, 2137–2144.
Nero, V., Farwell, A., Lister, A., Van Der Kraak, G., Lee, L. E. J., Van Meer, T., Dixon, D. G. (2006). Gill and liver
histopathological changes in yellow perch (Perca flavescens) and goldfish (Carassius auratus) exposed to oil sands
process-affected water. Ecotoxicology and Environmental Safety, 63(3), 365–377. doi:
10.1016/j.ecoenv.2005.04.01410.1016/j.ecoenv.2005.04.014
Kumari, U., Mittal, S. & Mittal, A.K. (2012). Surface ultrastructure of the gill filaments and the secondary lamellae
of the catfish, Rita rita, and the carp, Cirrhinus mrigala. Microsc. Res. Tech. 75: 433–440
Dar B A, Qureshi T A and Khaliq R, 2011. Effect of Cadmium Chloride on the Histoarchitecture of Certain Organs
of a Freshwater Catfish, Clarias batrachus. Our Nature, 9: 83-88
Kole RK, Banerjee H. and Bhattacharyya A. (2001). Monitoring of market fish samples for Endosulfan and
Hexachlorocyclohexane residues in and around Calcutta. Bull Envirron Contam Toxicol 67: 554–559
Kaoud, H. A., Zaki, M.M., El-Dahshan, A. R., Saeid, S. and EL Zorba, H. Y., 2011. Amelioration the toxic effects of
cadmium-exposure in nile tilapia (Oreochromis Niloticus) by using Lemnagibba L. Life Sci. J., 8: 185-195
Hanna, M.I., Shaheed, I.B. and Elias, N.S., 2005. A contribution on chromium and lead toxicity in cultured
Oreochromis niloticus. Egyptian J. aquat. Biol. Fish., 9: 177-209.