Chapter No.

01

GENERAL INTRODUCTION

Toxicology is the study of harmful and unfavourable effects that chemicals or physical agents
have on living things. The "study of poisons" is what it actually is. It provides details on how a
toxicant component is conveyed to humans as well as aquatic life. The adverse consequences can
take many different forms, ranging from instant death to subtle alterations that are seen months
or years later. They may be invisible at several levels of the body, such as an organ, a kind of
cell, or certain molecules.

Toxicologists are regarded as toxicology's founders. Many chemicals, according to Parcelsus,

were carefully considered for their potential damage to plants or animals. He also observed the
body's response to those substances, which depended on how much of each chemical was
consumed in order to determine if the drug was dangerous or helpful.

Toxic or poisonous substances could be organ toxins or systematic poisons. If a toxic substance
influences the whole body or numerous body parts instead of specific location is said to be
systematic poison. If a toxic substance affects the specific part or organ of body, it is said to be
target organ toxicant. The mechanical advancement is possibly the main cause for the
development of contaminated zones that are, for sure, releasing the environment pollutants.
Particularly careless use of pesticides is a global problem that is being examined across the globe
(Cimbaluk et al., 2018). Toxicology is the scientific study of how chemicals can have harmful
effects on living organisms, most notably in the environment. Through a variety of pathways,
heavy metals are introduced into the maritime environment and have a fatal impact on marine
life's physiology and morphology.

Approximately (hours/days) after exposure, severe toxic effect occurs. It often consists of one
dosage or a combination of dosages taken during a period of 24 hours. Chronic poisoning
exposure occurs repeatedly for a several weeks or months before becoming dangerous. Chronic
toxicity which causes entire destruction to certain organ systems and might take months or years
to manifest as clinical disorders.

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Development toxicity quality is related to harmful toxic organisms either directly by affecting
marine life or indirectly by disrupting the ecological food chain (Ganesan & Thirumavalavan).

The primary factors that affect fish physiology, reproduction, development, and behaviour are
the characteristics of the water (Z. U. Shah & Parveen).

The main criteria for human and aquatic life existence may be clean water accessibility. The
existence of clean water is essential to life. Within the ecosystem of estuaries, lakes,
wetlands, coves, seas, rivers, and groundwater, several types of water are available. However,
humans have a big impact on them, and that has an impact on aquatic life. Climate and
geographic variations are a few natural factors that affect the quality of water ( P. S. Kumar,&
Rathi, 2021). Rapid industry, population growth, and urbanisation have all contributed to serious
water contamination. The release of unprocessed, clean and toxic mechanical effluent is one of
the primary causes of freshwater pollution. The harmful contaminants discharged into sewage
systems can also harm marine life (Vardhan, Kumar, & Panda 2019).

Pesticides are mostly transported by water from one application area to other areas of the
ecosystem. When a living thing enters an aquatic environment, it undergoes a variety of changes,
including ones to its pace of development, food, behavioral habits, and so forth.

For many aquatic life forms, natural poisons including pesticides, metals and organics pose real
risks. Therefore, an extraordinary amount of research has been done to determine how toxicants
affect the physiology and ability of many aquatic organisms to survive( Scott & Sloman, 2004).
Water body pollution has rapidly increased around the world and has been recorded in several
places. Since fish serve as a crucial link in the food chain and are contaminated with pesticides, it
is vital to learn more about the harmful effects of toxins, especially pesticides. Fish are a vital
protein source and play a role in human growth (Jugino Mankuzhiyil Sivanandan & toxicology ,
2021)

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Rivers offer an abundant source of fresh water to living beings, rendering them essential for
survival. Despite comprising just 0.0001% of the Earth's overall water proportion, rivers convey
water and essential nutrients to the biosphere (Anhwangeb et al., 2012).

As rivers are perceived as convenient disposal sites for wastewater in developing countries, the
deterioration of water quality in river systems has emerged as a major environmental
apprehension for these nations in recent times (Jamil N et al., 2015).

Pakistan has four primary installations on the Chenab River:

Marala Headworks - situated close to Sialkot, Khanki Headworks - located in Gujranwala
District, Qadirabad Headworks - situated in Mandi Bahauddin District, and Trimmu Barrage -
located in Jhang District. The significance of the Chenab River, Pakistan's second-largest river,
lies in its vital role for irrigation, livestock, and aquaculture purposes (Bhatti M.T et al., 2011).
The concentration and dispersion of specific organochlorine pesticides in sediment samples
extracted from the River Chenab, Pakistan, were analyzed. The Chenab's catchment area
encompasses the most fertile agricultural zone in Pakistan. Unregulated utilization of agricultural
chemicals places significant strain on the water quality of the Chenab (Eqani S et al., 2011).
Moreover, the river traverses through industrialized and densely inhabited urban centers of
Pakistan, resulting in substantial discharge of anthropogenic wastewater into the river channel.
The initial gauging location is situated at Marala Barrage (74.46° E, 32.67° N), near Sialkot,
Pakistan. The Chenab River catchment area spans between 73°–78° E and 32°–35° N (Ahmed et
al., 2020).
The Chenab River catchment encompasses approximately 26,000 km2 of land until reaching
Marala Barrage. The elevation within the catchment fluctuates from 235 m near Marala Barrage
to 7103 m in the higher snowy region (Shahzad, A et al, 2018, 11, 383). The gradient of the
Chenab River ranges from 25 m/km in its uppermost section to 0.4 m/km in the flat plains
(Awan, S.A et al., 2003).
A thorough investigation is necessary to understand the hydrological flow patterns in the Chenab
River. Since the chemical composition of rivers is influenced by complex interplays between
various physical, chemical, and biological factors, exploring river basins provides valuable

3
understanding of the hydrological processes involving the diverse elements within the catchment
(Brezonik and Arnold 2011; Stumm and Morgan 2012).
Within Pakistan, the contamination of aquatic environments can occur through the deposition of
air-borne pollutants and the direct release of waste containing PCBs into rivers, lakes, and
coastal areas. Both these pathways serve as potential sources of pollution in aquatic ecosystems
(Biterna and Voutsa, 2005; Eqani et al., 2012a; Eqani et al., 2012b).
In the developing countries of South Asia, such as Pakistan, Nepal, India, and Bangladesh, where
urban areas intersect with rivers and their tributaries, the discharge of industrial waste, domestic
sewage, and agricultural runoffs results in substantial pollution levels (Karn and Harada, 2001;
Kambole, 2003; Pekey et al., 2004; Richardson et al., 2015).
The environment comprises five primary elements: soil, water, climate, indigenous flora, and
land formations. Among these, water holds utmost significance for human existence. Moreover,
it plays a vital role in the sustenance of various other ecosystems (K. S. Adu-Manu et al., 2017).
Rivers play a vital and fragile role in supporting the survival of every living species. The
preservation of these freshwater ecosystems is at risk due to the declining condition of their
watercourses. Rivers across the globe serve as the main sources of water for residential,
industrial, and agricultural purposes (Jain, 2009).
The presence of perilous chemicals in industrial wastewater (effluents) exerts a substantial
influence on the contamination of aquatic environments, altering the physical, chemical, and
biological composition of the recipient water system (Eqani et al., 2012). Specifically, the
unregulated release of larger quantities of industrial and municipal wastewater is contributing to
a gradual rise in the presence of toxic metal pollutants within the river ecology of Pakistan
(Jabeen et al., 2012).
As a result of their transfer to aquatic organisms and subsequent incorporation into the food web,
heightened concentrations of trace metals in aquatic sediments can pose a risk to human well-
being (Salati and Moore, 2010).
In the Florida Keys, maintaining a pristine water quality is imperative for the well-being of the
marine ecosystem. Seagrass and coral reef populations flourish in clear, nutrient-poor waters.
However, excessive levels of nitrogen in the water can trigger the proliferation of harmful algae,
leading to the destruction of seagrass and corals. Various pollutants, including metals, oils,

4
pesticides, and fertilizers, enter the water from the surrounding land, exacerbating the growth of
excessive algae and causing other detrimental impacts. The quality of river water holds immense
significance as these water sources serve multiple purposes. They provide drinking water for
households and businesses, support agricultural irrigation, facilitate hydroelectric power
generation, contribute to transportation and infrastructure systems, foster tourism and recreation,
and fulfill various other human and economic water-related needs (Venkatramanan et al,2014).
Water, being the second most essential element for the existence of life following air, is
thoroughly elucidated in scientific literature regarding its quality. The phrase commonly
employed to delineate water quality is "the characteristics pertaining to the physical, chemical,
and biological aspects of water."(Spellman, 2013).
The evaluation of water condition is conducted with regards to the needs of one or multiple
living organisms and/or any human necessity or objective in order to ascertain its quality (Shah,
2017). Three separate indicators of water quality include: Physical indicators, Chemical
indicators, and Biological indicators. Turbidity refers to the level of haziness in water and is
measured by the velocity at which light travels through it. The primary factors contributing to
turbidity are clay, silt, organic matter, plankton, and other suspended particles in the water (E.R.
Alley, 2007).
The expenses associated with water treatment for different purposes are increasing. The capacity
of harmful bacteria to hide within particles acts as a protective shield, rendering them resistant to
the disinfection process (Edzwald, 2010).
Harmful chemical particles have the potential to damage fish gills, leading to a reduction in their
disease resistance as well (Kiprono, 2017).
Heavy metals such as cadmium, lead, and mercury, along with other harmful organic pollutants
like pesticides, polychlorinated biphenyls (PCBs), and polycyclic aromatic hydrocarbons
(PAHs), have the ability to be efficiently absorbed by suspended particles. An increase in
turbidity leads to elevated water temperatures, which, in turn, reduces the availability of food
since suspended particles absorb more solar heat. Furthermore, warm water holds less dissolved
oxygen compared to cold water, resulting in a potential decrease in the concentration of
dissolved oxygen (DO). To measure turbidity, a nephelometric turbidimeter is utilized,

5
converting it into units known as Nephelometric Turbidity Units (NTU) or Turbidity Units (TU).
One TU is equivalent to milligrams of suspended silica per milliliter. The human eye cannot
perceive turbidity levels above NTU, although turbidity in heavily contaminated water can
approach NTU. Groundwater often exhibits relatively low turbidity due to natural filtration
processes that occur as water seeps through the soil (Viessman W et al. 2004)
Temperature has an influence on various characteristics such as taste, consistency, solubility,
odors, and chemical reactions. Consequently, temperature plays a role in sedimentation,
chlorination, and the measurement of biological oxygen demand (BOD). Moreover, the
biosorption of dissolved heavy metals in water is also influenced by temperature. Generally,
people prefer water within the temperature range of 15 to 30 degrees Celsius (Abbas SH, 2014).
The temperature of water holds great significance for the well-being of all aquatic species.
Temperature significantly influences both biological and chemical processes within aquaculture
operations. It stands as one of the foremost external factors affecting fish production. Deviations
from the optimal temperature range hinder fish growth, and extreme temperatures can even lead
to fatalities (Joseph and associates, 1993).
Water obtains its hue from the degradation of organic substances, particularly plants, as well as
inorganic materials such as dirt, stones, and rocks. This discoloration is generally considered
undesirable more for aesthetic purposes rather than health concerns. The color of a water sample
can be determined by comparing it with reference colors or using colored glass discs. The color
produced by a platinum solution (potassium chloroplatinate, KPtCl) at a concentration of 1 mg/L
is considered equivalent to one unit of color. The color of a water sample can be described as
follows: The overall color of the water sample, encompassing both dissolved and suspended
components, is referred to as the apparent color. Once the water sample has been filtered to
eliminate any suspended matter, the true color can be determined. The number of color units can
range from 0 (clear) to 70. Pure water has zero color units as it is colorless (APHA.2005).
The presence of foreign substances in water, such as dissolved gases, inorganic compounds, or
organic components, can contribute to alterations in taste and odor. These sources can originate
from various origins, including natural sources, domestic activities, or agricultural practices
(DeZuane J.1997).

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When evaluating water quality, one of the primary factors to consider is its pH level. By utilizing
a simple pH sensor or test kit, the acidity or alkalinity of the water can be determined. Acidic
water is characterized by a higher concentration of hydrogen ions, whereas basic water contains
more hydroxyl ions. The pH of water is a critical parameter and is defined as the negative
logarithm of the hydrogen ion concentration (Spellman FR. 2017).
The pH scale encompasses a range from 0 to 14, where a value of 7.0 represents neutral water.
Readings below 7.0 indicate acidity, while readings above 7.0 indicate alkalinity. Pure water has
a pH value of 7.0. Rainfall, on the other hand, typically has a pH of 5.6, indicating a slightly
acidic nature. Drinking water within the pH range of 6.5 to 8.5 is considered safe. Shifting pH
levels can have various effects on plants and animals, including:

1. Aquatic plants and animals have specific pH requirements for their survival, and even slight
changes in pH can negatively impact their quality of life. Slightly acidic water can harm fish
membranes, irritate their gills, and reduce the hatching of fish eggs. Aquatic plants and animals
can perish when exposed to water with excessively high pH levels. Low pH levels can be fatal to
frogs due to their skin's sensitivity to pollutants.

2. The productivity of a reservoir is significantly influenced by the quantity of hydrogen ions

present, which is determined by pH levels. Optimum pH levels for fish growth typically range
from 6.4 to 8.3 (Robert et al., 1940).
The pH level in fish cage culture can undergo a decline due to the accumulation of waste
deposits (Demir et al., 2001). In the course of the summer, Jiwyam and Chareontesprasit
discovered a higher pH (7.8 to 8.8) in Halali reservoir (Jiwyam and Chareontesprasit ,2001).
Elevated photosynthetic activity and the breakdown of external organic matter within the lake
could be attributed to this phenomenon, as heightened temperatures lead to increased nutrient
levels. Additionally, the inflow of sewage and agricultural waste can contribute to a rise in water
pH. Processes such as food degradation and respiration produce carbon dioxide, which interacts

7
with water to create carbonic acid and hydrogen ions, thereby acidifying the environment
(Mallasen et al., 2012).
The presence of acids in a solution is a measure of its acidity. "Acidity" refers to the quantitative
capacity of water to neutralize a strong base to a specific pH level. Water acidity is primarily
caused by mineral acids, carbon dioxide, and hydrolyzed salts such as ferric and aluminum
sulfates. Acids can impact various processes, including biological activity, chemical reactions,
and corrosion. Acidity arises when carbonic acid (H2CO3), derived from the dissolution of
carbon dioxide in water, is present. This can originate from the atmosphere or as a result of
respiratory processes in aquatic organisms. The level of acidity is determined through titration
using standard sodium hydroxide (0.02N) and phenolphthalein as an indicator (APHA.2005).
The alkalinity of water indicates its ability to counterbalance acids. Evaluating the alkalinity of a
water sample is commonly done to determine the appropriate amount of soda and lime to add for
water softening purposes. Water softening is particularly beneficial in preventing corrosion in
boilers. The presence of bicarbonate, carbonate, and hydroxide ions contributes to an increase in
water alkalinity. If water tests indicate high levels of alkalinity or acidity, it is likely that the
water has been contaminated in some way. Low-alkalinity water has less buffering capacity
compared to high-alkalinity water. Bicarbonates also serve as a storage system for carbon
dioxide, allowing for increased carbon dioxide production during photosynthesis. This, in turn,
ensures a continuous supply of oxygen to the system. Alkalinity values in freshwater systems are
typically within the range of 5 to 500 mg/L (Lawson, 1995).
To promote fish production in fertilized ponds, it is recommended to maintain a total alkalinity
level of at least 20 mg/L (Boyd 1982).
Changes in the overall concentration of alkalinity are primarily influenced by bicarbonates. Total
alkalinity can be utilized to assess the productivity and state of water bodies (Jiwyam and
Chareontesprasit, 2001). The water in the Halali reservoir exhibited abundant nutrients and had
an alkalinity level ranging from 90 to 160 mg/L (Tamot et al., 2008).
The alkalinity of water in Indian reservoirs ranges from 40 to 240 mg/L (Sugunan,2011). The
alkalinity level in Nile tilapia cage culture measured 156.75 to 191.16 mg/L (Jiwyam,2012).

8
Alkalinity values exceeding 20 mg/L were deemed beneficial for the productivity of tilapia
(Lucas and Southgate, 2012).
Naturally, groundwater, streams, and lakes possess a certain amount of chloride, but when
freshwater exhibits relatively elevated chloride concentrations (250 mg/L or higher), it may
indicate contamination from wastewater. Chlorides can find their way into surface water from
various sources, including wastewater, agricultural runoff, and rocks containing chlorides. While
chloride ions (Cl) in drinking water do not pose any immediate health risks to the general
population, excessive amounts can impart a salty taste. Chlorides are generally not harmful to
individuals, although the sodium content in table salt has been linked to kidney and
cardiovascular issues (Chatterjee A.2001).
Water exhibits hardness when it contains a significant quantity of minerals. If left unattended,
dissolved minerals in water can deposit scale on hot water pipes. When showering in water with
a high mineral content, you may find it challenging to generate a lather with the soap you are
using. The hardness of water is primarily attributed to the presence of magnesium and calcium
ions, which can enter water through soil and rocks. Generally, groundwater tends to have higher
water hardness compared to surface water. The hardness of water can be determined using a
colorimeter or test strip. The hardness of water is determined by the concentration of calcium and
magnesium salts, which are primarily combined with bicarbonates and carbonates to produce
temporary hardness, and sulphates, chlorides, and other mineral anions to produce permanent
hardness, respectively. One degree of hardness corresponds to 17 ppm CaCO3 (Jiwyam and
Chareontesprasit, 2002).
Soft water, characterized by a calcium carbonate (CaCO3) concentration of 0 to 75 ppm,
possesses minimal buffering capacity. On the other hand, extremely hard water, with a CaCO3
concentration exceeding 300 ppm, exhibits the highest buffering capacity. Hard water falls
within the range of 150 to 300 ppm CaCO3 concentration (Boyd, 1990; 1998).
In contrast to the wet and cold seasons, summer exhibits an elevated level of overall hardness
(Hujare, 2008).

9
 The increase in water hardness can be attributed to the decrease in water volume and the
escalation of evaporation rate during elevated temperatures (Thirupathaiah et al., 2012).
A crucial measure of water quality used to assess the contamination level of rivers, lakes, and
streams is the quantity of oxygen dissolved in the water. The presence of a significant amount of
dissolved oxygen indicates excellent water quality. Dissolved oxygen is a result of the solubility
of oxygen in the water. In aquaponic systems and other aquaculture facilities, dissolved oxygen
(DO) plays a vital role in water quality. Fish obtain oxygen through passive diffusion across their
gills from the water. To facilitate this passive diffusion, a substantial concentration of dissolved
oxygen in the water is required, enabling the oxygen to move from the water into the
bloodstream, driven by a concentration gradient (Colt et al,2002).
When dissolved oxygen (DO) concentrations fall below the necessary levels, fish are unable to
efficiently convert energy into a usable form. This detrimentally affects their growth rate, feed
efficiency, and swimming ability (Jones and D.R. 1971).
Fish react to reduced dissolved oxygen (DO) levels by accelerating their opercular respiration
and exhibiting gasping behavior (Wedemeyer and G.A 1996).
To ensure the well-being of both cold water and warm water fish species, it is recommended to
maintain dissolved oxygen (DO) levels at a minimum of 6 parts per million (ppm) for cold water
fish and 4 ppm for warm water fish. This limit is suggested to safeguard the health of the fish
(Wedemeyer and G.A 1996). Due to its representation of the physical and biological processes
occurring in water, dissolved oxygen serves as a vital parameter for assessing water quality. A
reservoir with a dissolved oxygen (DO) concentration of 5 mg/L is advantageous for year-round
fish culture (Amankwa ah et al., 2014).
The level of dissolved oxygen (DO) serves as an indicator for the degree of contamination in
aquatic environments. It plays a crucial role in supporting and maintaining life, as well as
facilitating the decomposition and degradation of organic substances. The rainy season tends to
exhibit a wider range of dissolved oxygen levels due to heightened wind activity and the mixing

10
of monsoon rainfall. DO has garnered significant attention as a water quality indicator in relation
to eutrophication levels. The content of dissolved oxygen in water is affected by factors such as
temperature, dissolved salts, wind velocity, pollution load, photosynthetic activity, and
respiration rate (Tamot et al., 2008).
Biological oxygen demand (BOD) is the requirement for oxygen by bacteria and other
microorganisms as they consume organic matter, which serves as their nourishment during the
metabolic process (Tchobanoglous G et al,2003).
If this process occurs in water, the dissolved oxygen within a water sample will be depleted.
When there is a significant amount of organic matter present, a considerable amount of dissolved
oxygen is consumed to facilitate the decomposition of the organic matter. However, this poses a
challenge as aquatic life requires dissolved oxygen for survival. The biological oxygen demand
(BOD) can be determined using the dilution method, providing insight into water toxicity. The
BOD serves as an important indicator of the pollution level in a body of water. BOD refers to the
fraction of dissolved organic matter that is rapidly and efficiently digested by bacteria. It
quantitatively measures the presence of degradable organic matter that consumes and depletes
the dissolved oxygen in water. Higher BOD levels lead to unpleasant and unhealthy
environments characterized by foul odors. The growth of BOD levels can be attributed to the
favorable environmental conditions for microbial activities at elevated temperatures (Tamot et
al., 2008).
Higher values of biochemical oxygen demand (BOD) are directly indicative of the pollution
level and exhibit an inverse relationship with dissolved oxygen (DO) concentration. This is
because BOD is directly linked to the decomposition of deceased organic matter within the lake.
BOD concentrations in the Hathaikheda reservoir near Bhopal varied from 0.0 to 4.0 mg/L
(Namdev et al., 2011).
The indication of water quality can be greatly determined by the presence or absence of living
organisms. Biologists have the ability to assess water quality by observing the presence of fish
and insects in natural water bodies and utilizing a species diversity index (SDI) (Nathanson
JA.2004)
Bacteria are single-celled microorganisms that can thrive and reproduce rapidly under favorable
conditions of optimal pH, food availability, and water temperature. Due to their exponential
growth, it is challenging to accurately determine the quantity of bacteria in a water sample. In

11
cooler water, bacteria generally multiply at a slower rate. Elevated levels of bacteria in water can
pose a risk of various waterborne infections, including cholera, tularemia, and typhoid. Algae,
consisting of small photosynthetic organisms, utilize pigments to convert inorganic substances
into organic matter through the utilization of solar energy. This process involves the absorption
of carbon dioxide and release of oxygen. Algae play a vital role in wastewater treatment methods
employing stabilization ponds. Synthetic pesticides are categorized into organophosphates,
organochlorines, and carbonates. Organophosphates, in particular, have been extensively used in
agriculture and are highly effective against pests. Chemicals introduced into the environment
through industrial effluents, household waste, and agricultural runoff can harm water sources,
which are crucial for all forms of life. The continuous discharge of agricultural wastewater into
water bodies often leads to the accumulation of pollutants, increasing the risk of hazardous
contamination. The ongoing use of toxic substances contributes to the ongoing deterioration of
the environment (Vander Oost, R. et al.,2003).
Pesticides can enter the environment through various pathways, including direct and indirect
applications. Significant quantities of pesticides are directly applied to agricultural fields,
primarily aimed at pest control and enhancing crop yields. Over time, these chemical substances
find their way into water reservoirs, resulting in the destruction of aquatic plants and animals.
Such contaminants pose a potential threat to non-target organisms, including fish (Pandey et
al.,2015).
Fish in direct contact with the aquatic environment are susceptible to any modifications that may
arise within it. Moreover, adjustments in fish osmoregulation, carbohydrate metabolism, overall
body physiology, hematological characteristics, and reproductive functions are anticipated
(Shreshth ,et al.2013).
Exposure to toxins results in alterations in size, characteristics, membrane fragility, and
lysosome stability. Extensive research has demonstrated the ability of purified alkaline
phosphatase to degrade various types of polyphosphates, including synthetic phosphate,
inorganic pyrophosphates, and metaphosphatase with an average chain length. Multiple team
members have observed abnormalities in fish gills (Akhtarunnessa and colleagues, 2012).
As urban wastewaters are significantly more concentrated compared to rural streams, the
processes of urbanization, agricultural fertilizer usage, and various other human activities can

12
alter the chemical composition of rivers. This effect is particularly pronounced in industries such
as mining and the chemical sector (Meybeck et al., 1989; Meybeck, 1996;
Flintrop et al., 1996).
Furthermore, novel sources such as reservoirs, which expedite evaporation and capture calcite
and silica, as well as irrigated soils with inadequate drainage that can retain soluble elements,
contribute to the alteration of water quality. To address these human-induced impacts, various
measures have been implemented, including mining and industrial practices, urbanization,
population density rates, fertilization and irrigation practices, and the construction and operation
of reservoirs. Despite the potential effectiveness of wastewater treatment and recycling in
reducing significant ions originating from mining operations (such as those involved in
petroleum and gas extraction, coal and lignite mining, pyritic ores, potash, and salt) as well as
industries and other sources, the comprehensive evaluation of their impacts remains scarce.
In most instances, urban wastewater treatment has minimal effects on the main ions. The
influence of human activities on silica is still not well comprehended, particularly at typical pH
levels found in surface waters. There is currently no conclusive evidence indicating a substantial
surplus of silica in rivers due to urban or industrial pollutants. The gradual alteration of river
chemistry has been observed early on, as exemplified by the systems of the Mississippi and Saint
Lawrence rivers concerning SO42- ions (Berner, 1971).
According to regularly conducted studies, the levels of sodium (Na+), chloride (Cl), and sulfate
(SO42-) have been consistently increasing worldwide since the 1960s. On the other hand, the
concentrations of calcium (Ca2+), magnesium (Mg2+), and bicarbonate (HCO3-) have remained
relatively stable (Kimstach et al., 1998).
Certain rivers (such as the Rhine, Weser, Vistula, and Don) that undergo the effects of mining
might experience more profound alterations compared to rivers affected solely by development
and industrialization (like the Mississippi, Volga, and Seine). The concentration of ionic
substances gradually increases when river water is redirected and employed for agricultural
purposes, particularly sodium (Na+), potassium (K+), chloride (Cl), and sulfate (SO42), as
observed in the case of the Colorado, Murray, and Amu Darya rivers. Agriculture can also
contribute to the process of salinization, similar to what occurred with the Neman River.
Assessing the various sources of significant ions caused by human activities, which vary
depending on the factors mentioned above, poses a challenge. Furthermore, these human factors

13
change over time within specific societies and reflect diverse environmental concerns, giving rise
to various types of interactions between rivers and communities (Meybeck, 2002).
Gaining knowledge about the diversity of life forms is essential for grasping the workings and
operations of ecosystems. In this period of environmental transformation, sustaining ecological
processes relies heavily on comprehending patterns of variety in the natural world (Pecl et al.,
2017).
The Kingdom Fungi carries out unique functions in both land and water ecosystems through the
breakdown of organic substances and engaging in symbiotic or parasitic relationships with other
organisms, including animals and plants (Peay et al., 2016; Hawksworth and Lücking, 2017;
Grossart et al., 2019).
In recent studies, scientists have been investigating the variety of fungi found on different
surfaces in land-based and water-based environments, all thanks to the growing adoption of
DNA metabarcoding methods (Duarte et al., 2015; Peay et al., 2016; Nilsson et al., 2019). On the
contrary, fungi demonstrate notable regional variation and geographic diversity (Bahram et al.,
2015, 2016).
Studying the expansive range of fungal diversity, encompassing vast spatial scales such as
landscapes spanning tens of kilometers and regional scales spanning hundreds to thousands of
kilometers, demands significant investment in labor, analytical resources, and financial means.
Recent investigations have explored the potential of assessing biodiversity by analyzing
environmental DNA present in river water (Deiner et al., 2016; Nakagawa et al. 2018).
The water of a river has the ability to carry diverse data for various eukaryotes in both water-
based and land-based environments by merging genetic material from the surrounding region
(covering a distance of several tens of kilometers) (Deiner et al., 2016).
It is expected that spores and mycelia of land-based fungi could penetrate rivers, facilitating the
identification of fungi from both terrestrial and aquatic environments (Voronin, 2014).
Normally, waterways are composed of a system of multiple tributaries that diverge from the
primary stream. Geographically proximate regions exhibit similar fungal populations, which
mirror the surrounding habitat and the dispersal abilities of individual species within terrestrial
fungal communities (Talbot et al., 2014; Matsuoka et al., 2016b; Peay et al., 2016).
If the makeup of the microbial communities on the surrounding land is mirrored in the fungal
DNA compositions found in the river water, the DNA assemblages also reveal the spatial

14
arrangement. The flow of water in rivers is unidirectional, moving from upstream to
downstream. Consequently, the DNA assemblages of river water can unveil distinct structures
exclusive to that particular river. The resemblance between samples from the same branch and
those from different branches may be more pronounced. Studying the vast geographical diversity
of fungi necessitates an understanding of fungal DNA assemblages in river water and their
spatial organization. Nonetheless, there has been no previous investigation into the geographic
configuration of fungal DNA assemblages in river water.
In this study, our objective is to examine and elucidate the spatial structure of fungal DNA
assemblages in river water, encompassing branches across forested landscapes. We specifically
aim to determine (1) if fungal DNA assemblages exhibit greater similarity between river
branches compared to within branches and (2) whether spatial patterns in fungal DNA
assemblages are influenced by environmental or geographic distances.
Various estuarine and coastal environments have been documented to experience eutrophication,
which is characterized by the enrichment of nutrients leading to increased aquatic primary
productivity. This often manifests as the presence of harmful algal blooms and hypoxic bottom
water conditions (Officer et al., 1984; Rosenberg, 1985; Westernhagen et al., 1986; Benović et
al., 1987; Justić et al., 1987; Anderssonand Rydberg, 1988; Smayda, 1990; Cooper and Brush,
1991; Hickel et al., 1993; Turner and
Rabalais, 1994; Justić et al., 1995a; Rabalais et al., 1996; Rabalais et al., 1999).

Fish

Fish are a particular kind of aquatic life. Fish are the only animals with gills for breathing. The
majority of fish have scales to protect them, fins to aid in swimming, and streamlined bodies to
facilitate movement through the water. Fish are frequently utilised as sentinel species for
toxicological studies because they may quickly consume dangerous compounds and can react
more strongly to them at high concentrations. This is because fish play distinct functions in the
trophic web. (Cimbaluk et al., 2018)

Fish are aquatic in nature and cold blooded animals that move and balance themselves with the
aid of their fins and breathe via their pharyngeal gills. 40 percent of the known fish species—or
over 22,000—live in freshwater and river waters. Such fish are primarily found in tropical

15
waters. According to reports, India is home to around 2500 species, with about 930 of those
being freshwater species.

A great source of protein is fish. They have a large selection of both items and by products. For
many fishermen and locals in the study region, fishing is their primary source of income.
Numerous researchers have examined the fish flora of diverse bodies of water, according to a
survey of the literature (Parimala, 2021).

Labeo rohita (Rohu)

In rivers all around Asia, the Labeo rohita (Rohu) freshwater carp is most common in India,
Pakistan, Bangladesh, Nepal, Myanmar, and Thailand. They have an important economic role
and generate 1.6 metric tonnes of meat annually.

Production of rohu is mostly done for processing and consumption. The rohu for fish sausage's
chemical and physical characteristics were on par with those of jarred meat sauces. According to
reports 30% or more of the rohu generated in a natural processing facility was discarded as trash
with little or no market value, using the skin to produce a high-value product is the consequence.
In particular collagen is an option that is known to boost sales for the company while lowering
disposal costs (Savedboworn, Kithiphattanaboworn, Benjakul, Sinthusamran & Kishimura,
2017).

Rohu is widely distributed and may be found in ponds, lakes, and rivers. It is a bottom feeder and
a herbivore. It often feeds on algae and aquatic plants. It often uses its air bladder to suck down
air and breaths via its gills. In July and August, it reproduces in moving water. It is oviparous,
and water bodies serve as the external site of fertilization.

Its body has a silvery sheen on the ventro-lateral sides and a blackish dorsal color. It weighs
between 20 and 25 kg and has a length of 1 meter. The body is made up of the tail, the trunk, and
the head. Its body is formed like a spindle. Except for the head, its entire body is covered with
scales. It is a precious species that is grown extensively and in high demand in Asian countries.
For the purpose of tracking growth variability, survivorship, the suitability of pond conditions,
and the creation of fish processing machinery, study into the morphometric dimensions of rohu
carp is essential. (Jayraj, Machavaram, Sahu & Paradkar, 2019).

16
It’s a schematic figure of Labeo rohita that highlights the various body sections. About one
metre long, compressed, fusiform body. the body is silvery white on the bottom and blue or
brownish on the back.

The body is covered with cycloid scales which are massively overlapped. Because of the sunken
skull, the nose is short, oblique, and blunt. A pair of filamentous barbs encircles the upper lip. A
definite lateral line may be seen. There are soft fin rays, paired pectoral and anal fins, and dorsal
fins.

Split by two equal lobes, the caudal fin. Between the bladder and the inner ear is where you'll
find the weberian apparatus. A mesonephric organ is the kidney.

For a sizable group of customers who live close to freshwater resources, especially in developing
nations, fish serves as both a source of high-quality protein and additional cash. Pakistan's per
capita fish consumption is fairly low compared to the global average at 2 kg, although demand is
constantly rising. Fish flesh is thought to be a superior and affordable source of poly unsaturated
fatty acids of the omega series and has greater water content than red meat while having a higher
test saturated fat level. Fish is a rich source of the necessary vitamins A, B, and D as well as the
amino acids methionine, cysteine, lysine, threonine, and tryptophan. The evaluation of fish flesh
components is required for consumer education and the creation of consumer applications. The
suitability of fish feed as a source of protein and the nutritional value and amino acid statuses of
fish for processing (Hussain, Sultana, Ahmed & Mahboob, 2018).

17
Adults occupy the river. They are a diurnal species that are often solitary. They occasionally
tunnel. Feeding plants is also necessary. The southwest monsoon typically coincides with the
time of year when fish spawn. In a river that is flooded, spawning occurs. The fecundity of fish,
which ranges from 226,000 to 2,794,000, is determined by the fish's length, mass, and ovary
weight. spawns much above tidal extends in rivers' middle sections when flood water spreads in
largely lipid shallows over fertile flats. moreover, bundh-type tanks and storage tanks.

When they are young, rohu favors the zooplankton they consume, which is mainly composed up
of cladocerans and rotifers, with phytoplankton serving as a backup food supply. All
zooplanktonic organisms are significantly positively chosen for feeding during the fingerling
stage, along with certain smaller phytoplankton such as phytoflagellates desmids and algal
spores. However, most of phytoplankton is strongly positively selected in adults. In its juvenile
and adult stages, Rohu is primarily a herbivorous column feeder, favoring algae but also
vegetation that is submerged. Further evidence that it feeds the ground comes from the presence
of decomposed organic matter, sand, and mud in its gut. Adults do not exhibit the same
schooling behaviour as juveniles do.

Evaluation of fish digestive tracts provide crucial information on feeding behaviours, including
qualitative and quantitative assessments of eating patterns. Information about eating patterns is
helpful for making decisions about natural resources. An essential part of fish populations is the
study of fish diet and feeding behaviours.Fish diet combines a variety of ecologically significant
elements, including as calorie intake, inter- and intraspecific relationships, behavioural condition,
and biodiversity utilization. It is possible to evaluate prey choices using food, eating patterns,
and gut content, Impacts of development and reproduction strategies (M & Biosciences, 2020).
The most prized of all fish species raised in traditional or modern aquaculture systems in India is
the Labeo rohita. The Godavari and Krishna rivers, as well as other locations of India outside of
its natural range, have all been used for fish farming. It has also been brought to many other
countries, particularly the old Soviet Union, Japan, as well as and the Philippines, because of its
fast growth and good meat.

18
Notopterus Notopterus

A member of the Notopteridae family, Notopterus (Pallas, 1769) is found in India, Bangladesh,
Pakistan, and other parts of Asia. It lives in stagnant and slow-moving waters in ponds, lakes,
floodplains, and canals. However, in captivity, it is raised in tanks throughout much of India. For
reproduction throughout the mating season, it migrates from freshwater to brackish waters (Riehl
and Baensch, 1989). It consumes tiny fish, crustaceans, insects, and even the young root of
aquatic vegetation. It migrates to where it spawns on rainy days during its yearly breeding season
before returning to its regular home during the dry season (Rainboth, 1996).

This fish is commercially captured using a variety of small-scale fishing gear and is highly
important economically as a food fish in Borneo and Sumatra in Southeast South Asia. It is a
popular fish for human food and is enjoyed in both fresh and dried forms. Its soup is beneficial
for measles patients (Roberts, 1992). Threats to its existence have not been assessed, despite the
fact that it is important as food.

Rita rita

In big rivers all throughout the Indian subcontinent, particularly in the Ganges and Indus River
Drainage, Myanmar is home to the bagrid catfish Rita rita (Hamilton, 1822) of the genus Rita
Bleeker, 1854. This native fish may grow to a maximum size of 50 cm and a weight of 2 kg in
Pakistan, where it is found in Balochistan, KP, Punjab, and Sindh (Rafique and Najam Ul Huda
2012). . It is preferable to have only large, powerful fish in its tank since, while being a rather
calm fish, it will consume lesser tank mates. Large cyprinids, especially Labeo spp., make good
tank companions.However, information on the rita rita's body composition and breeding has not
yet been published.

Typically, the majority of a fish's live weight is made up of 2-12% fats,0-30%proteins and 70–
80% water. (Love, 1970). However, the values vary significantly within every species and are
also based on things like size, sexual preference, diet, season, exercise, etc. The biochemical
makeup of fish is of great importance for determining their dietary preferences and assessing
their physiological requirements at various stages of development. Therefore, proximate fish
components are crucial for assessing the species' nutritional value and physiological state.

19
(Gershamovich et al., 1984; Brown and Murphy, 1991). In addition to being a good source of
protein, fish is also rich in other nutrients including iodine, lipids, vitamins, and minerals.
Understanding the fish's functional characteristics is crucial for using it in the creation of
products with value-added. The examination of the water, fat, protein, and ash content of fish is
known as proximate body composition. Because they are present in trace amounts and are
typically disregarded in normal analyses, carbohydrates and non-protein substances (Cui and
Wootton, 1988). The amount of water in anything is an excellent predictor of how much protein,
fat, and energy it contains. (Dempson, et al., 2004).

Pesticides are artificial or natural compounds used to attract, entice, pulverize, or control pests.
(Lushchak, Marvishyn, Haak, Story, & Storey, 2018). Pesticides can enter the body in a number
of ways, depending on the species, metabolic characteristics, and reactions to toxin (Lushchak et
al. 2018)

Pesticides are among the many anthropogenic substances that damage surface waterways across
the world. Pesticides mostly reach the aquatic environment by runoff from agricultural regions,
effluents from waste water treatment facilities, and spray runoff and water from agricultural
areas or unintentional spills. Pesticide concentrations in surface waterways are often at their
highest following application on agricultural or private areas, and frequently during heavy rain
events due to storm water runoff and seepage. When such runoff comes from rural regions, it
typically has a large amount of suspended particles, which causes pesticides and soil fragments
to be transported simultaneously into surrounding water bodies(Knauer, Homazava, Junghans,
Wemer, & management, 2017).

In any event, the pesticides that are originally employed to attack the target species have a higher
poison quality and less target specificity, which are damaging to other creatures. According to
investigations, several insect sprays and fungicides are toxic to some kinds of aquatic life,
including fish. Environmental risks evaluation of pesticide risk must be done in order to meet the
administrative prerequisites.

A crucial organic indicator of the pollutant's ecological threat is its acute toxicity to fish (Jia, liu,
Wang and chemistry, 2020) Modification Hood components are essential bio monitoring tools
used in toxicological research to quickly assess a compound's invertebrate toxicity. Oceanic life

20
forms' blood reflects any unfavorable changes in water quality by separating the blood cells with
slender epithelial layer.

Exogenous factors like strain continuously cause significant changes in blood composition. For
instance, variations in the quantities of hemoglobin (Hb), white blood cells (WBC), red blood
cells (RBC), and other essential components occurred in response to physiological and
biological situations. Extreme molecular level changes in organisms caused by the introduction
of pesticides are thought to be the most sensitive and immediate instances of any damaging
toxin. The most dangerous substances have an effect on living things at their most basic level by
producing compounds or metabolites in response (Juginu Mankuzhiyil Sivanandan &
Toxicology, 2021).

Question about the use of wild to agricultural pesticides flares among the many forms of waste
from fish farms. Since angle feed and angle oil were replaced with plant-based raw materials in
fish bolster, the current bolster is made up of 70% plant fixes (Cho, 20201). Testing has revealed
that modern day supports used in fish farming may contain additional levels of chemicals.
(Olsvik et al., 2019). A pesticide can be any substance or mixture of substances intended to
prevent the destruction, repelling, or lessen the harm caused by any bug. It might include creepy
crawlies, plant pests, avian, warm-blooded, and other species that interact with one another to
feed, destroy habitats, grow, or provide help as well as organisms that transmit and spread
illnesses and are viewed as a nuisance. The most often used pesticides include rodenticides,
herbicides, and sprays for insects and other pests (Yadav, Devi, & engineering, 2017).

Depending on the situation, there are many different forms of pesticides. Pesticides that are
maintained by plants and animals and may spread to untreated tissues are known as systemic
pesticides. Some pests control chemicals are also linked to other species and migrate through
them. Non-systemic pesticides are a subset of pesticides that kill by touch rather than ingestion,
however they nonetheless affect the targets they were intended for. The most popular and
effective method of classifying pesticides according to their chemical composition or type of
dynamic foxings. This classification offers details on the adogacy, external features, and internal
chemical makeup of certain weed killers (Yadav et al., 2017). Incorrect use of pesticide may
have been considered the extreme solution to pest issues that recurred in the 1940s and 1950s,
but for many unexpected and unanticipated istopes First off, gradual exposure of a species to a

21
single chemical has undermined the foundations of sustainable populations, especially with
regard to cyclodiones. Second, because to heavy pesticide use, the small pests have evolved into
the big pests, outpacing both their natural enemies and themselves. Third, pesticides have a
severe adverse effect on non-target living things, especially those that reside in the oceanic sinks
of poisons, where any substance given mechanically is sure to end up sooner or later (Munty,
2018).

Classification based on the original pesticide's ingredients may include chemical and natural
compounds that aim to eliminate harmful agents or prevent harm brought on by pests. Based on
the source of root pesticide, a pesticide might be classed as chemical or biological. The most
convenient natural weed herbicides to use are those with specificity. While synthetic pesticides
are typically used on a large scale that inflicts a great deal of non-target life forms, the act at
targeting pests is unmistakably tied to live beings. Bio-pesticides are often ecologically friendly
because they need little usage and are less potently deterred. Chemical pesticides heavily
contaminate the environment since they are highly prickly and never fully biodegrade (Yadav et
al, 2017). Several instances of fish deaths in large numbers that occurred right away after
pesticides were heavily applied have been documented. Fish deaths that occurred simultaneously
in 15 tributaries of the Terese Steam near Albama in 1950 were described. These fish deaths
followed the use of a chlorinated hydrocarbon pesticide and were linked to precipitation nudging
away from rural regions (Murty, 2018).

Pesticides can infiltrate seagoing systems in a variety of ways, for example, after being applied
to crops, where they may be washed from drainage systems and into aquatic habitats if it rains. It
is crucial to consider the potential genotoxic effects of creepy crawly executioner on creatures
other than pests. Organophosphates have been used as insecticides for centuries, but they are also
dangerous as alkylating agents and potential genotoxicants. The populations of these from larger
distances have expanded in China and Indo-Pakistan, two Asian countries. However, negative
effects on microbes, invertebrates, fishes, and land- and water-based creatures can be particularly
severe in sea-going biological systems (Garcés, Pires, Rodrigues, & Health, 2020).

22
 Literature Review

Saleem (1983) analyzed the biological, chemical, and hydrological characteristics of three
waterways within Nottingham. The limnology of freshwater bodies in Libya was explored by
Khan (1989) and Zarmouth (1989). Kulshresther et al. (1992) identified the limnological traits of
India's Mansarover reservoir, while Unni et al. (1992) examined the physico-chemical and
biological parameters of the Narmda River during that same year. In 1996, Imbert and Standford
conducted an ecological investigation on a regulated prairie watercourse situated in Western
Montana.

In Denmark, Kjeldsen (1996) carried out field trials to examine the impact of phosphorus, light
intensity, and grazing by invertebrates on the control of algal biomass in a small lowland stream.
Karr (1998) employed the IBI method, an innovative approach utilizing multiple metrics, to
investigate the river biology and provide insights for landscape management. Meanwhile,
Hubble and Harper (2002) scrutinized the structure and progression of the phytoplankton
community within the water column of Lake Naivasha in Kenya. Rothhaupt (2000) conducted
research on the fluctuations of planktonic communities in Germany. Similarly, in Lake Sempach,
Switzerland, Biirgi and Stadelmann (2002) examined alterations in the composition and diversity
of phytoplankton prior to and following a restoration process.

Kagalou et al. (2002) undertook a study on the water quality of the Kalamas River in Greece.
DeBruyn et al. (2003) examined the impact of sewage on a significant river food web in Canada.
Williams et al. (2003), Morselli et al. (2004), Gevrey et al. (2004), and Angermeier and
Davideanu (2004) investigated water quality-related concerns in various streams and rivers.

23
Singh et al. (2004) conducted research in India to analyze the effects of toxicants from treated
and untreated wastewater discharged by sewage treatment plants on agricultural productivity,
environmental quality, and human health.

Javed and Hayat (1999) conducted research on the harmful effects of heavy metal toxicity within
the aquatic ecosystem of the Ravi River. Javed and Mahmood (2000) documented the
accumulation of metals in the organs and tissues of fish from the same river. Farid et al. (2003)
reported the contamination of city effluent from Faisalabad with heavy metals. Ali et al. (2004)
examined the monthly fluctuations in the physico-chemical properties and metal concentrations
of the Indus River at Ghazi Ghat, Muzaffar Gharh, Pakistan. In a publication by Sarda, P., and
Sadgir, P. (2015), an assessment of various indicators was conducted to evaluate the quality of
surface water bodies. The parameters considered encompassed physical, chemical, and biological
aspects such as water temperature, acidity (pH), electrical conductivity (EC), total dissolved
solids (TDS), dissolved oxygen (DO), chemical oxygen demand (COD), biochemical oxygen
demand (BOO), chloride content, alkalinity, overall hardness, calcium and magnesium levels,
sulphate concentration, nitrate content, ammonia nitrogen, nitrate nitrogen, and nitrite nitrogen.
The objective of the research was to pinpoint the key comprehensive water quality indicators
derived from surface water bodies that are crucial for drinking, irrigation, and aquatic
ecosystems. Several scholars deliberated on various facets related to water quality. Surface water
bodies encompass a range of extensive and straightforward field parameters, such as water
temperature, acidity (pH), electrical conductivity (EC), total dissolved solids (TDS), dissolved
oxygen (DO), chemical oxygen demand (COD), biochemical oxygen demand (BOD), and other
factors, all of which hold equal significance for diverse purposes.

Sunanda, M., et al. (2016) conducted a review highlighting that among the many harmful
substances ever released into the environment, pesticides pose a significant threat. While
pesticides have contributed greatly to improving human well-being, they also have considerable
adverse effects on non-target species. Various hazardous chemicals, including those found in
urban or industrial areas, pose a direct risk to freshwater organisms, especially delicate species
like fish, through runoff and leaching into groundwater. Chlorpyrifos, an organophosphate
pesticide, poses an exceptionally high level of danger to freshwater fish. Given the importance of

24
fish in the food chain, studying the impact of pesticides such as chlorpyrifos on fish can provide
insights into their potential harm to humans.

Murthy, K. S., et al. (2013), emphasized the heightened risk posed by pesticides employed for
pest control in agricultural regions, particularly for non-target organisms such as fish. These
chemicals can adversely impact the health of fish by impeding their metabolic processes and, in
severe cases, leading to fatalities. The present research aims to examine the potential detrimental
effects of pollutants and pesticides on fish, with the acquired data potentially assisting in the
assessment of environmental hazards faced by both freshwater and marine organisms.

Gorde et al. (2013) investigated the pivotal role of water in shaping the Earth's surface and
regulating climatic conditions. Among the myriad of molecules, water stands out as one of the
most vital substances profoundly impacting human existence. The physical, chemical, and
biological attributes of water are frequently employed to assess its quality. Unfortunately, due to
rapid industrialization and the unregulated usage of chemical fertilizers and pesticides in
agriculture, water quality is deteriorating, leading to the depletion of aquatic organisms. The
consumption of contaminated water exposes humans to waterborne diseases, emphasizing the
need for regular water quality monitoring. Key parameters such as temperature, acidity (pH),
cloudiness (turbidity), salinity, nitrates, and phosphates can be measured to assess water quality.
Additionally, surveys of aquatic macroinvertebrates offer valuable insights into water quality
conditions.

Ullah, S., and Zorriezahra, M. J. (2015) delved into the field of ecotoxicology, providing a
comprehensive review of the toxicity induced in fish by pesticides. The agricultural sector
globally relies heavily on pesticides to enhance crop productivity while minimizing labor and
effort. However, fish, among other non-target organisms, are susceptible to harm from pesticide
exposure. High doses of pesticides often lead to immediate mortality, whereas lower, sub-lethal
concentrations can cause a multitude of detrimental effects with potentially fatal consequences.
These alterations may manifest in the behavior of the exposed fish, encompassing variations in

25
feeding patterns, aggression or avoidance responses, and reproductive behavior. Additionally,
they can manifest in other forms, such as modifications in the fish's blood components (red blood
cells, platelets), organ structure (liver, kidneys, gills, muscles, brain, and intestine), antioxidant
defense mechanisms (glutathione reductase), or overall physiology. Enzymes like peroxidase,
catalase, superoxide dismutase, glutathione peroxidase, and glutathione-S-transferase may
induce changes in the nutritional composition (proteins, lipids) as well as impact carbohydrates,
moisture content, ash content, fish quality, and even hormonal or enzymatic fluctuations
(genotoxicity). Several environmental organizations are actively involved in addressing this
issue, which relates to the prevalence of banned substances. Despite their prohibited status, these
compounds continue to be accessible for purchase. The continuous production and widespread
use of newly manufactured pesticides (such as insecticides and herbicides) contribute to the
ongoing deterioration of conditions for aquatic organisms, with fish being particularly affected.

In their study, Malik et al. (2020) conducted a review focusing on the examination of diverse
environmental factors, highlighting the significance of water in conjunction with temperature,
oxygen levels, alkalinity, and photoperiod for developmental processes. Consequently,
conducting research and assessments to evaluate the influence of fish nutrition and water quality
becomes essential for enhancing fish growth and size.

Fatima et al. (2020), however, assert that this represents the utmost essential requirement.
Unfortunately, human activities have resulted in its contamination, posing hazards to aquatic and
terrestrial life. Heavy metals stand out as one of the most notable pollutants. These heavy metals
can accumulate within fish, presenting health risks when they are present in the water.
Urbanization and industrial activities have contributed to the pollution of the aquatic
environment by these heavy metals, disrupting the balance of the ecosystem and impacting
aquatic organisms. Among the most prevalent contaminants are zinc (Zn), lead (Pb), and nickel
(Ni), with zinc also being a significant pollutant in sediment. Fish exposed to these metals can
develop a range of hazardous conditions, including deformities, metabolic issues, kidney disease,
liver ailments, cancer, infertility, and behavioral abnormalities. These pollutants indirectly affect
human health as fish consumption is a staple in the human diet.

Assessment of environmental risk is a crucial step in the licensing and approval process for
pesticides. Calculated exposure doses are used to determine a substance or formulation's

26
potential impact on aquatic populations (Knauer et al., 2017). Fish fatality studies are still
necessary to evaluate the environmental danger of both new and existing pollutants in the aquatic
compartment, and one of the main tasks of today's aquaculture environmental sciences is to
estimate concentration response evaluations (Rodrigues, Varela, Pardal, & Oliveira, 2019).

On earth, water is a vital resource for survival of life, but because of population increase,
industrialization, untrained exploitation of natural water resources, and haphazard urban
development, water quality has dramatically declined (Vardhan et al., 2019).

Aquatic species are presently threatened by the water contamination of the aquatic environment
caused by a variety of pesticides. Organophosphorus insecticides are used on crops including
rice, cotton, and vegetables to suppress large insects and mites. These substances pose a serious
threat to the aquatic ecology, especially to fish (Geetha & Economics, 2018).

Because of industrialization and urbanization, several contaminants are entering aquatic habitats
both directly and indirectly. It is common practice to utilize behavioral bioassays for toxicity
testing. While bioassays focused on reproduction and growth need lengthier bioassays, those
based on behavior are quicker, more sensitive, and more relevant to the environment. A system's
activity, reaction, or performance under particular set of conditions is characterized as behavior
and attitude, which has an influence at the organismal level. We believe that an improved
knowledge of cognitive responses during chemical exposure may be improved (Sharma, Thakur,
Verma, Sharma, & Studies, 2019).

Over stimulation and suppression of respiratory activity are two of the most crucial signs of
chemical pollution toxicity in aquatic medium. Oxygen utilization and ventilator pattern both
continue to decline steadily. The behaviour and physiology of living creatures are affected by
high pesticide doses in the environment, notably the respiratory system. The majority of
chemicals have a tendency to build up in the tissues of aquatic animals. It is quite challenging to
evaluate how pesticides affect aquatic creatures that are not their intended targets ( Kallagadda &
Venkatarathnamma ). Organic contaminants that are introduced to the aquatic ecosystem might
change in a number of ways. An important method for determining the ecosystem's surviving
structures and the toxicity of contaminants is biotransformation. For the protection of several
crops, the usage of such organophosphate pesticides (OPPs) has expanded. Aquatic ecosystems

27
can get contaminated by organophosphate in a number of ways, including spray drift, surface
runoff, and subsurface water leaching. As the amount of organophosphate that enters the aquatic
environment is unknown (Sumon, Rashid, Peeters, Bosma, & Van den Brink, 2018). An
extensive family of chemicals, pesticides containing organophosphorus exhibit a wide range of
physiochemical properties as well as considerable toxicological activity and effects. They are
primarily employed to manage pest populations in a range of food (fruits and vegetables, tea,
etc.) and non-food (tobacco, cotton, etc.) crops. Although malathion is a commonly used
organophosphate insecticide, nothing is known about how it affects fish. Because of this, the
study is set up to examine the effects of Malathion on rohu (Labeo rohita) carp in a semi-static
systems with a variety of characteristics. Histopathology of the liver showed a number of
modifications, including hepatic necrosis, fatty infiltration, hemorrhage, and vacuolation.
Swellings, fluctuating, glycogen, and congestion (Ullah et al.2018). In agriculture, the
cultivation of flowers, pest management, plastic production, flame retardants, and a variety of
domestic uses, organophosphate pesticides (OPs) are often utilized. Due to their relatively quick
rate of disintegration, OPs, which are phosphoric acid ester forms, are often recognized as safe
for agricultural usage. Variable toxicities can be caused by acute or long-term OP exposure in
people, animals, plants, and insects. These are said to decrease the function of acetyl
cholinesterase in insects, as well as terrestrial and aquatic creatures, leading to defects in the
respiratory system, neurological system, hepatic system, and kidney system. Organophosphates
interfere with the growth-promoting process by impeding a number of plant-growth-essential
enzymes, transcuticular diffusion, and permeability. Due to the environmental problems caused
by excessive amounts of organophosphates in meals and water sources, it appears that there is an
urgent need to create a rapid, dependable, and affordable technology that can be used on-site. We
examine how environmental toxins are classified, their environmental effects, assessment
methods, and eco-friendly bioremediation strategies for effective biological remediation (Sidhu
et al., 2019).

With over thirty-three thousand species, fish represent the most diverse group of vertebrates.
These may be distinguished by, among other visual characteristics, their shape, colour, and head.
It might be difficult for some people to distinguish between the various varieties of fish readily
available for purchase. Researchers have an interest in using machine vision algorithms to
categorize fish species based on photographs of their visual traits. However, the effectiveness of

28
the classifier is dependent on the caliber of the training data set. A dataset of drawings is needed
to study the classification algorithm's pattern. The data from fish-Pak is particularly useful for
evaluating various classifier parameters, such as activation functions, momentum, and their
influence on overall performance (S. Z. H. Shah et al., 2019). Large carp big food resources and
very high protein sources found in Indian big carp are frequently consumed by Labeo rohita as
part of numerous toxicity testing investigations in the biological sciences and medicine.
Numerous toxicology techniques developed in lower animals exposed to environmental toxins
like herbicides, pesticides, insecticides, heavy metals, etc., as well as behavioral, hematological,
enzyme, recovery, histopathological, immunological, and other studies may help researchers
concentrate on the important areas of research that have not yet been identified (Jayakumar,
Senthilmurugan, Vijayan, & Tamizhazhagan, 2018).

Any product used to eliminate or manage pests, such as insecticides, herbicides, and fungicides
(fungi), is referred to as a "pesticide" in the generic sense. One main supporter of the green
revolution sought to improve and employ safe weed killers to combat the vast array of insect and
herbal pests that have an adverse effect on the quantity & quality of food production worldwide.
This study investigates the definitions, categories, and toxicity of pesticides as well as the
environmental destiny, effects on human health, and strategies for detection, disposal, and
treatment (Hassaan & El Nemr, 2020). Fish oxygen intake is impacted by pesticide pollution
from a variety of classes, including organophosphates, carbonates, and other new generation
pesticides, at sub lethal quantities. As well as exhibiting signs of pesticide toxicity, respiratory
responses to dangerous levels increase breathing volume, suggesting that the influence on the
surface of the airway was significant in fish (Prasanna, Anithasmruthi, Venkatarathnamma, &
Toxicology, 2020). Pesticides are compounds or blends of substances with similar physical,
chemical, and biological characteristics. Therefore, these are grouped according to the traits.
Certain weed killers are certainly divided into various groups according on how they are used.
The three most widely used weed killer classifications today depend on the pesticide's mode of
action, the pest life forms it kills, and the consistency of its chemical makeup. Improper pesticide
application can have detrimental effects on both living things and their environment. The
majority of pesticides all kill pests and other relevant incidental living species without making a
distinction between them (Yadav et al., 2017). Three historical eras must be used to group
pesticide usage. Several terms, such as chemical categories, structure of molecules, biological

29
activities, and toxicity, are used to categorize pesticides. Because pesticides employ chemical
substances to kill pests and manage weeds, they may also be dangerous to air, water, soil, and
crops in addition to aquatic life such as birds, fish, and beneficial species. Additionally, pesticide
contamination extends outside of the actual plants, causing environmental damage. Due to their
contamination of the environment and food supply, these chemical residues have an effect on
human health. Additionally, the use of pesticides is influenced by climate change-related
variables, which results in increased control of weeds and chemical pollution (Tudi et al., 2021).
Due to their quick biodegradation and lack of persistence, organophosphate insecticides are
employed in agriculture to control pests, but their wide range of adverse effects extend well
beyond the pest. To investigate the virulent effects of pro enofos (50 EC) on blood parameters,
such as red blood bug pests that have a negative impact on the quantity & quality of global food
production, one of the key proponents of the green revolution undertook an experiment. This
research looks at how pesticides are classified, how dangerous they are, what they do to the
environment, how they affect people's health, and how to identify, get rid of, and treat them.
(Hassaan & El Nemr, 2020). Fish oxygen intake is impacted by pesticide pollution from a variety
of classes, including organophosphates, carbonates, and other new generation pesticides, at sub
lethal quantities. As well as exhibiting signs of pesticide toxicity, respiratory responses to
dangerous levels increase breathing volume, suggesting that the impact on the surface of the lung
was significant in fish (Prasanna, Anithasmruthi, Venkatarathnamma, & Toxicology, 2020).
Pesticides are compounds or blends of substances with similar physical, chemical, and biological
characteristics. Therefore, these are grouped according to the traits It is true that some herbicides
are categorized according to how they are used. The three most widely used weedkiller
classifications today depend on the pesticide's mode of action, the pest life form it kills, and the
consistency of its chemical makeup. Improper pesticide application can have detrimental effects
on both living things and their environment. The majority of pesticides all kill pests and other
relevant incidental living species without making a distinction between them (Yadav et al.,
2017). Three historical eras must be used to group pesticide usage. Several terms, such as
chemical categories, molecular structure, bioactivities, and toxicity, are used to categorize
pesticides. Chemicals are included in pesticides, which are used to kill pests and control weeds.
As a result, aquatic life, such as birds, fish, and beneficial species, as well as water, air, soil, and
crops, may be poisoned by pesticides. Additionally, pesticide contamination extends outside of

30
the actual plants, causing environmental damage. Due to their contamination of the environment
and food supply, these chemical residues have an effect on human health. Additionally, the use
of pesticides is influenced by climate change-related variables, which results in increased
suppression of weeds and chemical pollution (Tudi et al., 2021). Due to their quick
biodegradation and lack of persistence, organophosphate insecticides are employed in agriculture
to control pests, but the wide range of adverse effects extend well beyond the pest. A research
study looked at how proenofos (50 EC) affected blood-related factors such haemoglobin,
glucose, red blood cells (RBC), and white blood cells (WBC). Giemsa stained blood smears
showed erythrocytic cellular abnormalities (ECA) including acanthocytes, fusion expanding
shaped, tear-drop shaped, and twin when compared to the control. The frequency of MN
formation also increased significantly at various toxicant doses. The current investigation amply
revealed that profenofos does negatively affect the haematology of a carp. An effective pesticide
for both agriculture and non-agricultural applications is malathion. Mosquitoes, flies, domestic
pests, animal parasites (ectoparasites), and body and head lice have all been successfully
controlled using malathion. The pesticide has been proven to be extremely hazardous, even to
aquatic creatures that are not its intended targets, including fish. The objective of the study was
to ascertain how the pesticide affected various biochemical traits (Lekeshmanaswamy, 2018).
The effects of many chemicals on fish's haematological and blood metabolic processes are
summarised in the current appraisal of the literature from 2000 to 2019. Usually, the noticed
alterations point to anemia, inflammation, and hyperglycemia. Stress and intoxication are all
linked to hypoproteinemia, a rise in cortisol levels, and hepatic aminotransferase activity. Other
adjustments that could be seen, including a rise in red blood cell properties, signify a retaliatory
response. The signs of immunosuppression, which are frequently seen, are indications that
pesticides have a deleterious impact on the immune system and may lead to immunosuppression.
A number of variables, such as the active ingredient or its concentration, the period of exposure,
the fish species, environmental circumstances, and others, can have an impact on pesticide-
induced pathophysiological alteration in fish (Bojarski, Witeska, & Research, 2020). A typical
organophosphate molecule utilised as a fire-prevention additive in a variety of products is
triphenyl phosphate. However, its use has a detrimental effect on a number of species. At
ecologically relevant concentrations, only a few research have looked at the acute toxicity of
TPhP in aquatic species that are not targeted.The purpose of this study was to examine the

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effects of short-term TPhP exposure on antioxidant properties, oxidative stress, and biological
molecules conversion. Genetic equilibrium, chromosomal credibility, programmed death of cells,
and pathogenic changes were investigated in Labeo rohita fingerlings.

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Aims and objectives

The study “water quality and fish body of fish composition of river Chenab, Punjab, Pakistan”
was done to examine to study the fish body composition and effect of water quality of river
Chenab and the effect of toxicity on fish body. The main objectives of this study was

 To check the water quality of river Chenab, Punjab Pakistan.

 To determine the effect of water quality on the growth of fish.
 To check the body composition of fish found in river Chenab Punjab Pakistan

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