Review Article

Induced-Toxicity of Pesticides on Edible Freshwater Fishes in Pakistan: A Review

Yaseen1, Asad Ullah2*, Imad Khan2, Maryam Begum3, Sumbal Bibi3, Umber3, Namra3, Abbas Khan4, Shumaila Gul5 and Raheela Taj6

1Department of Zoology, Abdul Wali Khan University, Mardan 23200, Khyber Pakhtunkhwa, Pakistan; 2College of Veterinary Science and Animal Husbandry (CVS and AH), Abdul Wali Khan University, Mardan 23200, Khyber Pakhtunkhwa, Pakistan; 3Department of Zoology, Women University, Swabi, 23430, Khyber Pakhtunkhwa, Pakistan; 4Department of Zoology, Government College University Lahore, 54000, Lahore, Pakistan; 5Department of Chemical and Life Sciences, Qurtuba University of Science and Information Technology, Peshawar 25000, Khyber Pakhtunkhwa, Pakistan; 6Institute of Chemical Sciences (ICS), University of Peshawar, 25120, Khyber Pakhtunkhwa, Pakistan.

Received | October 23, 2023; Accepted | December 29, 2023; Published | February 15, 2024

*Correspondence | Asad Ullah, College of Veterinary Sciences and Animal Husbandry, Abdul Wali Khan University, Mardan 23200, Khyber Pakhtunkhwa, Pakistan; Email: [email protected]

Citation | Yaseen, A. Ullah, I. Khan, M. Begum, S. Bibi, Umber, Namra, A. Khan, S. Gul and R. Taj. 2024. Induced-toxicity of pesticides on edible freshwater fishes in Pakistan: A review. Sarhad Journal of Agriculture, 40(1): 195-212.

DOI | https://dx.doi.org/10.17582/journal.sja/2024/40.1.195.212

Keywords | Pesticide, Toxicity, Effects, Edible, Fish, Pakistan

Copyright: 2024 by the authors. Licensee ResearchersLinks Ltd, England, UK.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

Introduction

Industrial advancements and development of novel technologies are essential to achieve success and comfort in ongoing era, but these are causing deterioration of precious natural water resources on earth (Tahir et al., 2021). Increasing number of industries, modernization and urbanization eventually steered us to the contamination of our environment and left us to face the problem of environmental pollution. These pollutants include domestic wastes, industrial effluents, agro-chemicals, etc. that include different organic and inorganic substances, heavy metals such as cadmium, arsenic, and pesticides. An after effect of urbanization and industrialization is that the pollutants released from industries eventually reach the aquatic bodies where they get dissolved and cause water pollution (Ullah et al., 2014). Environmental pollution is an elaborate phenomenon having multiple aspects with partial or unidentifiable origins (Ullah et al., 2014). International Agencies reported that such chemical substances have high potential and a constant threat to aquatic life including fish and bioaccumulation of these pollutants eventually entre into human food chain and is the main cause of human health hazard (Ullah and Zorriehzahra, 2015).

In view of its consumption by human, industrial production, industrial irrigation and stabilization of biodiversity, fresh water is extremely valuable in terms of preserving life. However, these ecosystems are at risk of suffering biodiversity losses due to being vulnerable to environmental pollutants including pesticides (Geist, 2011; Pisa et al., 2015; El-Murr et al., 2015; Javed and Usmani, 2015).

Pesticides are chemical compounds that are used to get rid of pests including insects, rodents, fungi and unwanted plants, herbs, weeds etc. (Akashe et al., 2018). These chemical compounds are grouped into different classes such as insecticides, herbicides, rodenticides, fungicides etc. According to the chemical nature of pesticides; it is further divided into groups like carbamates, organochlorine, organophosphate, pyrethroids (widely used), and aliphatic fungicides, inorganic rodenticides, amide fungicides, ammonium herbicides. Pesticides are a big source of potential environmental hazards to birds, fish, and other animals as well as humans when they infiltrate food chain (Khan et al., 2012). An increased burden of chemicals can arise in environment due to non-biodegradability of some of these classes of pesticide chemicals (Mahboob et al., 2011). Effluents with pesticides resulted in a marked rise in the mortality rate, growth retardation and tissue damage in fish (Rana et al., 2011). Susceptibility of different fish species to these pesticides is different at different concentration. The changes in different body parts and systems of fish have been observed to be different than each other as well as in response to different pesticides (Ullah and Zorriehzahra, 2015).

Pakistan is an agricultural country with most of its agricultural land irrigated by canal systems from rivers and their tributaries. Main rivers are Indus, Kabul, Chenab, and Ravi. Freshwater ecosystems of Pakistan have rich biodiversity including a large number of edible fish species such as Labeo rohita, Cirrhinus mrigala, Cyprinus carpio, Catla catla. Agriculture run-off, sewage wastes, fall into the rivers causing aquatic pollution and adversely affect fish life, leading to various hematological, biochemical, histopathological changes in fish body which when consumed by human’s results in different types of disorders in body. This review discusses research over past decade regarding toxic effects of pesticides induced in edible freshwater fishes of Pakistan and some future considerations.

 

General introduction of commonly-used pesticides

(Modified from Yadav and Devi, 2017).

Organochlorine pesticides (OCPs)

Organochlorines pesticides are organic compound attached with five or more chlorine atoms. Organochlorines (OC) are widely used in health sector and agriculture as insecticides and these are non-biodegradable. These pesticides have a negative effect on insects’ nervous system which cause disruption of the insect nervous system causing convulsions, then paralysis and eventually cause death, slowly and gradually. Most common examples of these pesticides includes: DDT, endosulfan, aldrin, lindane, dieldrin and chlordane. Production of DDT is now prohibited throughout the globe.

Organophosphate pesticides (OPPs)

Organophosphate pesticides are multi-purpose broad spectrum pesticides, which are derivatives of phosphoric acid used to get rid of different types of pests and act when inhaled, ingested, or penetrate in skin, leading to stomach abnormalities, nervous system impairments in affected organisms. These are biodegradable pesticides, causing little environmental pollution and show delayed pest resistance. They are more toxic to animals as they have cholinesterase-inhibiting properties leads to excess accumulation of acetylcholine neurotransmitter across a synapse. This results in failure of nerve impulses to move across the synapse causing muscular cramps, paralysis and eventually death. Most common OPPs include malathion, diazinon, glyphosate, and parathion.

Carbamates (Carb)

The carbamates share similar structural makeup with organophosphates(OP), but carbamates are distinct in that they originate from carbamic acid. Carbamates work by interfering with transmission of nerve signals resulting in the death of the pest by poisoning. Under natural conditions, they are easily degraded with little environmental pollution. They are occasionally used as fumigants, contact poisons, and poisons for the stomach. Insecticides that contain carbamates include carbaryl, carbofuran, propoxur, and aminocarb.

Pyrethroids (PYR)

Synthetic pyrethroid insecticides are produced from natural pyrethrins. Compared to pyrethrins, they are more stable and have a longer residual impact in environment. These insecticides are only mildly hazardous to mammals and birds, but they are extremely toxic to fish and insects. Majority of synthetic pesticides are non-persistent and quickly break down when exposed to light. Pyrethroids are considered safe to use in food. Cypermethrin and Permethrin are examples of popular synthetic pyrethroid pesticides.

Pesticide-induced toxic effects on fish

Behavior: Different studies on pesticides over past decade have shown alterations in behavior of various fish species such as sluggish swimming movements, lethargy, faintness, and disruption in swimming ability which renders the fish more prone to predators, affect their feeding, orientation, and territory defense (Prashanth et al., 2011). Interruption in behavior of fish by pesticides makes the fish stressed and immunocompromised, making them susceptible to different kinds of pathogens and infections (Nwani et al., 2013). In freshwater fish Labeo rohita commonly known as Rohu , imidacloprid induced morphological and behavioral changes such as avoidance mechanisms, abrupt and sluggish swimming movement in all directions, occasional jumping and hitting on the walls of tank, rapid scale loss, mucous secretion, change in body color (Qadir and Iqbal, 2016), while organophosphates (profenofos, trizaophos) and carbamates (carbofuran, carbaryl) caused suffocation, lethargy, descending movement, irregular swimming, gulping before death (Mustafa et al., 2014) in the same fish. Also, fipronil resulted in shakings, twitching, dizziness, increased operculum movement, body curving, breathing troubles in Cyprinus carpio (common carp) (Ghaffar et al., 2018). There are many different studies confirming the effects of different pesticides on behavior of different freshwater fishes of Pakistan (Mahboob et al., 2015a; Ghaffar et al., 2020, 2021; Usman et al., 2020; Zulfiqar, 2020; Akram et al., 2022; Wang et al., 2022) mentioned in Table 2.

Hematology: Blood parameters were assessed as physiological indicators of animals exposed to stressful conditions such as the presence of toxic substances, because blood acts as a patho-physiological reflector of the whole body (Velisek et al., 2012). Various pesticides have been extensively studied to trace for hematological changes such as changes in number of RBCs and WBCs, thrombocytes, neutrophils, hemoglobin content, hematocrit values in different freshwater fish species of Pakistan such as in Cirrhinus mrigala (Indian carp) due to diazinon (Rauf and Arain, 2013; Haider and Rauf, 2014), Chlorfenapyr, dimethoate, and acetamiprid (Ghayyur et al., 2021), Labeo rohita (Rohu) exposed to acetamiprid (Alam et al., 2014), triazophos (Ghaffar et al., 2015a), butachlor, a chloroacetanilide herbicide (Ghaffar et al., 2015b), Chlorpyrifos (Ismail et al., 2018), Diafenthiuron (Riaz-ul-Haq et al., 2018), thiamethoxam (Ghaffar et al., 2020; Hussain et al., 2020), fipronil (Ghaffar et al., 2021), pyriproxifen (Naseem et al., 2022; Li et al., 2022), Ctenopharyngodon idella (Grass carp) to endosulfan (Ullah et al., 2017), Cyprinus carpio (Common carp) in response to fipronil and buprofezin (Qureshi et al., 2016), fipronil (Ghaffar et al., 2018),Oreochromis mossambicus (Mozambique tilapia) to Chlorpyrifos (Ghayyur et al., 2019), Tor putitora (Mahaseer) to cypermethrin (Bibi et al., 2014), Oncorhynchus mykiss (Rainbow Trout) to Chlorpyrifos (Ali et al., 2020), Aristichthys nobilis (Bighead Carp) to acetochlor (Mahmood et al., 2022), triclosan (Akram et al., 2022), Hypophthalmichthys nobilis (Bighead carp) to pendimethalin (Wang et al., 2022).

Histopathology: Histopathological changes have been traced in different organs such as brain, gills, liver, kidneys, intestines, blood and muscles of different edible freshwater fish species of Pakistan. Pesticides-induced changes in histopathology of fish include lamellar disorder, lesions, hyperplasia, congestion, epithelial stimulating, micro gliosis, hemorrhages, necrosis, discoloration, neuronal degeneration, karyorrhexis, hepatocellular hypertrophy, and accumulation of pesticides residues in muscles (Khan et al., 2018). The pesticide damage tissue, effect function of Kidney, centers in spleen, edema, and disruption of cardiac myofibers in heart have been reported over past decade from various freshwater fishes. Some of the lethal pesticides are cypermethrin (Khan et al., 2018), mixture of endosulfan and chlorpyrifos (Naz et al., 2019), thiamethoxam (Ghaffar et al., 2020), profenofos and carbofuran to Labeo rohita (Mahboob et al., 2014a), cypermethrin to Tor putitora (Ullah et al., 2015), fipronil plus buprofezin to Cyprinus carpio (Qureshi et al., 2016), DDT and HCHs to Cyprinus carpio, Tor putitora, Glyptothorax punjabensis, Orienus plagiostomus (Aamir et al., 2016), Chlorpyrifos to Oncorhynchus mykiss (Ali et al., 2020), acetachlor to Aristichthys nobilis (Mahmood et al., 2022), pendimethalin to Hypophthalmichthys nobilis (Wang et al., 2022), Lambda-cyhalothrin to Ctenopharyngodon idella (Niaz et al., 2022). Some other researchers have also reported the histopathological changes in edible freshwater fishes of Pakistan (Rana et al., 2011; Nasir et al., 2012; Eqani et al., 2013; Bibi et al., 2014; Mahboob et al., 2015; Jabeen et al., 2015; Ullah et al., 2017; Qadir and Iqbal, 2016; Robinson et al., 2016; Karim et al., 2016a, 2016b; Riaz et al., 2018; Hussain et al., 2020; Ghaffar et al., 2021; Naseem et al., 2022; Li et al., 2022) (Table 2).

Biochemical (Oxidative stress) variation

Exposure to pesticides induces various biochemical changes in fish body. Chlorpyrifos cause increased activity of antioxidant enzymes in freshwater fish Oncorhynchus mykiss (Ali et al., 2020). Activity of anti-oxidant enzymes such as superoxide dismutase, peroxidase, and glutathione S-transferase increases in liver, gills, kidneys, brain, muscles, and heart of Labeo rohita when treated with mixture of endosulfan and chlorpyrifos (Naz et al., 2019). Rise in quantity of oxidative stress biomarkers and decline in concentration of antioxidant enzymes occur when Labeo rohita is exposed to pyriproxifen (Li et al., 2022), Aristichthys nobilis to acetochlor (Mahmood et al., 2022), Hypophthalmichthys nobilis to pendimethalin (Wang et al., 2022).

Total protein content (TPC)

Various studies showed the changes in protein content in various tissues such as liver, gills, intestines, blood, and muscles of different edible freshwater fishes of Pakistan when exposed to different pesticides. Total protein content declined in Cyprinus carpio when exposed to Karate i.e. λ-Cyhalothrin, protein value recorded in different tissues, Muscles 20.77±1.21a, Brain 26.01±2.06a and Liver 27.84±1.46a (Bibi et al., 2014), and fipronil and buprofezin (Qureshi et al., 2016). Exposure of Labeo rohita to Diafenthiuron, total serum protein (P = 0.004), cholesterol (P = 0.033) and creatinine (P = 0.002) were significantly reduced (Riaz-ul-Haq et al., 2018). Catla catla (Major Carp) recorded heights protein content range, 18.59±0.04 then used 0.038mg/L profenofos, reduce protein level 12.76±0.04, Labeo rohita (Rohu) protein content 19.18±0.02, used 0.06mg/L profenofos, decreased protein level 12.71±0.04. Cirrhinus mrigala (Mrigal carp) 14.62±0.02 protein value in healthy fish, used 0.041mg/L profenofos, reduced total protein level 8.70±0.01. showed decline in total protein when exposed to profenofos, (Ghazala et al., 2019). Oreochrmois mossambicus (Mozambique tilapia), also showed a decline in total protein content when treated with organophosphates, pyrethroids, and herbicide (Naqvi et al., 2017), and Chlorpyrifos (Ghayyur et al., 2019). Oreochromis niloticus (Nile Tilapia) also show a significant decrease in total protein content when exposed to malathion, Chlorpyrifos, and λ-Cyhalothrin, significant decrease total protein level in different tissue such as in brain (19.33±0.58) and muscle tissues (27.02±0.57) (Amin et al., 2021) and malathion (Zulfiqar, 2020). Exposure of Ctenopharyngodon idella (Grass Carp) to mixture of endosulfan+chlorpyrifos and endosulfan+bifenthrin decrease protein content interval of time reported in different tissues such as Hepatic, control group protein content 3.95±0.56Ba, lowest total protein recorded 1.98±0.21Bc, Muscle highest protein 1.04±0.17Ba and lowest 0.18±0.01Bc, Gills highest protein 2.86±0.33Ba and lowest 1.46±0.15Bc, Cardiac highest protein 0.86±0.12Ba and lowest 0.03±0.01Bc etc. (Usman et al., 2020), Lambda-cyhalothrin (Niaz et al., 2022) and Aristichthys nobilis (Bighead carp), to acetochlor (Mahmood et al., 2022) lead to decline in total protein content in investigated fish tissues followed by different standard procedure for determination of protein contents in different tissues of the different fresh water species.

Acute toxicity- LC50

Acute toxicity may be measured as oral, dermal and inhalational acute toxicity. LC50 is the measure of acute inhalation toxicity. Acute toxicity is expressed by various changes in behavior and physical activities, even death. However, different pesticides have distinct LC50 values in different organisms, determine by using Probit analysis and comparison was done by the APHA method (Mahboob et al., 2015). A list of common pesticides together with their lethal concentrations for different fish spe­cies have been listed in Table 1. For further information, “handbook of acute toxicity of chemicals to fish and other aquatic vertebrates” is also helpful (John­son and Finley, 1980).

Inhibition of acetylcholinesterase (AChE)

Organophosphates and carbamates apparently share the common mechanism of acetylcholinesterase inhibition at nerve endings that results in excess acetylcholine accumulation in the nerve ending overstimulating the effector organ. Studies have shown that acetylcholinesterase inhibition in fish has been associated with exposure of fish to different pesticide. In a study conducted by (Haider and Rauf, 2014), acetylcholinesterase activity was inhibited in Cirrhinus mrigala exposed to diazinon. Oncorhynchus mykiss also showed a decline in AChE activity when exposed to Chlorpyrifos (Ali et al., 2020). Inhibition of AChE activity occurred in Labeo rohita when exposed to profenofos and carbofuran (Mahboob et al., 2014), and Cyprinus carpio when treated with λ-Cyhalothrin (Bibi et al., 2014).

Genotoxicity

Genotoxicity is a property possessed by some substances that makes them harmful to the genetic information contained in organisms. Heavy metal ions and polycyclic hydrocarbons are the most influencing genotoxicants for fishes. Pesticides-induced genotoxicity has been reported in various freshwater fishes of Pakistan. Genotoxic threats and increased DNA damage have been noticed in Labeo rohita when exposed to triazophos, used standard blood test, staining method, serum enzyme such asALT, AST, ALP and measured by spectrophotometrically (Ghaffar et al., 2015a),endosulfan (Ullah et al., 2017), thiamethoxam (Hussain et al., 2020), and pyriproxifen (Li et al., 2022), Aristichthys nobilis when treated with acetachlor (Mahmood et al., 2022), and triclosan (Akram et al., 2022) and Hypophthalmichthys nobilis to pendimethalin (Wang et al., 2022).

 

Table 1: List of common pesticides along with their lethal concentrations for different fish species.

Pesticide	Experimental fish	LC50 value	Exposure time	Reference
Diazinon	Cirrhinus mrigala	8.15 mg L-1	96 hr	Rauf and Arain, 2013
Triazophos	Labeo rohita	6.64 mg L-1	96 hr	Mustafa et al., 2014
Profenophos	Labeo rohita	0.32 mg L-1	96 hr	Mustafa et al., 2014
Carbofuran	Labeo rohita	1.4 mg L-1	96 hr	Mustafa et al., 2014
Carbaryl	Labeo rohita	8.24 mg L-1	96 hr	Mustafa et al., 2014
Profenofos	Labeo rohita	0.31 mg L-1	96 hr	Mahboob et al., 2014
Carbofuran	Labeo rohita	1.39 mg L-1	96 hr	Mahboob et al., 2014
Karate (λ-Cyhalothrin)	Cyprinus carpio	0.16 µL L-1	96 hr	Bibi et al., 2014
Imidacloprid	Labeo rohita	550 mg L-1	96 hr	Qadir et al., 2015
Cypermethrin	Tor putitora	63 µL L-1	96 hr	Ullah et al., 2015
Triazophos	Cirrhinus mrigala	1.05 mg L-1	96 hr	Mahboob et al., 2015
Profenophos	Cirrhinus mrigala	0.21 mg L-1	96 hr	Mahboob et al., 2015
Carbofuran	Cirrhinus mrigala	0.49 mg L-1	96 hr	Mahboob et al., 2015
Carbaryl	Cirrhinus mrigala	4.75 mg L-1	96 hr	Mahboob et al., 2015
Fipronil	Cyprinus carpio	0.665 mg L-1	96 hr	Qureshi et al., 2016
Chlorpyrifos	Labeo rohita	442.8 µg L-1	96 hr	Ismail et al., 2014; Ismail et al., 2018
Endosulfan+chlorpyrifos	Oreochromis niloticus	5.64 μg L-1	96 hr	Ambreen and Javed, 2018
Endosulfan+chlorpyrifos	Labeo rohita	1.95 μgL-1	96 hr	Naz et al., 2019
Endosulfan+bifenthrin	Ctenopharyngodon idella	4.23 μg L-1	96 hr	Ambreen and Javed, 2015
Endosulfan+chlorpyrifos	Ctenopharyngodon idella	4.60 μg L-1	96 hr	Ambreen and Javed, 2015

 

Table 2: Effects of pesticide induced toxicity in different edible freshwater fishes of Pakistan studies over past decade.

S. No	Fish			Common name		Name of pesticide		Sample		Changes noticed	Reference
1	Labeo rohita			Rohu		Organochlorine and nitrogen-containing pesticides (endosulphan. DDE, parathion methyl, isoproturon, atrizine, carbofuran, carbaryl, deltamethrin)		Fish muscles		Accumulation of pesticides residues in fish muscles	(Rana et al., 2011)
2	Labeo rohita			Rohu		Imidacloprid		Live fish		Avoidance Mechanisms, sluggish and abrupt swimming movements in all directions, occasional jumping, hitting on the tank walls, rapid scale loss, mucous secretion, body color changed to light brown. 96 hrs LC50 value is 550 mg L-1	(Qadir et al., 2016)
3	Otolithes ruber			Tiger-toothed croaker		Organochlorines (HCH, DDT, dieldrin, eldrin)		Fish tissue		Bioaccumulation of Heptachlor exo-epoxide and Methoxychlor in Fishes	(Nasir et al., 2012)
4	Cirrhinus mrigala			Mrigal carp		Diazinon		Blood		Hematological alterations, Decrease in total RBCs, WBCs, Hb, and Hct values, LC50 value is 8.15mg/L, Increase in death rate with increase in diazinon concentration and exposure.	(Rauf and Arain, 2013)
5	Cyprinus carpio, Cirrhinus mrigala, Catla catla, Cirrhinus reba, Labeo calbasu,			Common Carp, Mrigal carp, Major Carp, Reba Carp, Orange fin labeo		Organochlorine Pesticides (OCPs) and Polychlorinated Biphenyls (PCBs)		Fish muscles		Intake of fish pose health hazard to humans	(Eqani et al., 2013)
6	Cirrhinus mrigala			Mrigal carp,		Diazinon		Blood		Decline in RBCs, WBCs, Hb, Hct, mean corpuscular	(Haider and Rauf, 2014)
7	Labeo rohita			Rohu		Profenofos, Carbofuran		Brain, gills, muscles, kidneys, liver, blood		Disturbed metabolism and neurotransmission, Profenfos toxicity is high compared to carbofuran in context of Acetylcholinesterase and Butyryl-cholinesterase inhibition in all tested organs	(Mahboob et al., 2014a)
8	Catla catla				Major Carp	Triazophos,		Live fingerlings		Acute toxics stress, 100% mortality at 2.8 mg L-1 carbofuran dose for 96 hrs, Behavior: suffocation, movement towards bottom, erratic swimming, lethargy, gulping before mortality	(Mahboob et al., 2014b)
						Profenofos,
						Carbofuran,
						Carbaryl
9	Catla catla				Major Carp	DDT, DDE, Endosulphan, Endosulfan sulphate, Cartap, Carbofuran		Flesh		Accumulation of pesticide in fish muscles	(Akhtar et al., 2014)
10	Cyprinus carpio				Common Carp	Karate (λ-Cyhalothrin)		Brain, liver, muscle tissue		Decrease in total protein content, Reduction of Acetylcholinesterase activity	(Bibi et al., 2014)
11	Labeo rohita				Rohu	Acetamiprid		Blood		Decrease in serum calcium, phosphate, and albumin, Increase in urea	(Alam et al., 2014)
12	Labeo rohita				Rohu	Organophosphates (Profenofos, Trizaophos) and Carbamates (Carbofuran, Carbaryl)		Live fish		Acute toxic stress, Behavioral stress exhibited by fish, suffocation, lethargy, fish rest at the bottom, irregular swimming, movement towards bottom, gulping before mortality.	(Mustafa et al., 2014)
										Table continued on next page.................
S. No	Fish			Common name		Name of pesticide	Sample		Changes noticed		Reference
13	Labeo rohita				Rohu	Triazophos	Blood		Microcytic hypochromic anemia, Decrease in lymphocyte and monocyte values, Increase in leukocyte count, DNA damage		(Ghaffar et al., 2015a)
14	Ctenopharyngodon Idella			Grass Carp		Endosulfan	Blood,liver, intestines, gills,		Blood: decline in total RBCs, WBCs, platelets, Hb, and PCV, Liver: degenerative changes, vacuolation, pyknosis etc. Gills: fusion and disruption, Intestines: atrophy of villi, increased goblet cells count		(Ullah et al., 2017)
15	Tor putitora			Mahseer		Cypermethrin	Blood, liver, gills, brain		Blood cells: decrease in RBSs count, Increase in WBCs count, Liver: glycogen vacuolization, hemorrhage vacuolization, congestion, fatty infilteration, hepatic necrosis, Gills : cellular infilteration, congestion, swollen tip of gill filament, hetropilic infilteration, damaged gill, Brain : discoloration, neuronal degeneration, Infilteration, severe spongiosis,		(Ullah et al., 2015)
16	Cirrhinus mrigala			Indian Carp		Triazophos, carbofuran, carbaryl, profenofos	Live fingerlings		Suffocation, Restlessness, Erratic swimming, Loss of equilibrium, Jerky movement, Mouth opened with rapid operculum movement, Lethargy, Gulping before death, 100% mortality at 1.6 mg L-1 of carbofuran for 96 hrs.		(Mahboob et al., 2015a)
17	Catla catla			Major Carp		Endosulfan, carbofuran, cypermethrin, profenophos, triazophos, deltamethrin	Fish muscles		Accumulation of endosulfan and profenofos in fish tissues, Higher concentrations of endosulfan, carbofuran and deltamethrin than permissible limits for fish set by International agencies, Health hazard to human and aquatic organisms		(Mahboob et al., 2015b)
18	Tor putitora			Mahseer		Cypermethrin	Liver, brain, gills		Liver: glycogen vacuolization, hemorrhage vacuolization, congestion, fatty infiltration, necrosis, Brain: discoloration, neuronal degeneration, infiltration and severe spongiosis, Gills: congestion, swollen tip of gill filament, cellular and hetrophilic infiltration, gill damage		(Ullah et al., 2015)
19	Labeo rohita, Channa marulius, Cyprinus carpio			Rohu, Great Snakehead, Common Carp		Pyrethroids, carbamates, neonicotenoids	Fish Muscles		Carbofuran detected in Labeo rohita, and Cirrhinus marulius, Cypermethrin detected in Cirrhinus marulius, Deltamethrin detected in Cyprinus carpio, and Cirrhinus marulius.		(Jabeen et al.,2015)
20	Labeo rohita			Rohu		Butachlor (Chloroacetanilide herbicide)	Blood		Decrease in RBCs, Hb, Hct, and lymphocyte value, Increase in total WBCs count, Morphological and nuclear abnormalities, mutagenic effects		Ghaffar et al., 2015b)
21	Hypophthalmichthys molitrix			Silver Carp		Deltamethrin	Liver, blood		Liver: necrosis, nuclear pyknosis, hypertrophy of cells, vacuolization, congestion of blood vessels, nuclear atrophy, Blood: Increased level of hepatic enzymes AST and ALT		(Karim et al., 2016a)
										Table continued on next page.................
S. No	Fish			Common name		Name of pesticide	Sample		Changes noticed		Reference
22	Hypophthalmichthys ii. Molitrix			Silver carp		Deltamethrin	Spleen, Kidneys		Kidneys: tissue damage, renal tubules degeneration,		(Karim et al., 2016b)
									Dilation, lyses and degeneration of glomerular capillaries, narrowing of the tubular lumen, atrophy, Spleen: tissue damage, necrosis, alterations in number of melano-macrophage centres
23	Cyprinus carpio			Common carp		Fipronil, buprofezin	Blood, Fish tissue		Decrease in total proteins content, globulin, RBCs, Thrombocytes, Hb, HCT, blood DNA content, Increase in WBCs and glucose concentrations, albumin unaltered, RBCs: necrosis, micro-nuclear formation and hyper-chromatosis, Gills: epithelial uplifting, lamellar necrosis, disorganization, fusion and atrophy, disruption of cartilaginous core, telangiectasia, Liver: hypertrophy of hepatocytes and nuclei, karyorrhexis, melano-macrophage aggregations, contractions of central vein, Kidneys: glomerular deterioration, Bowman’s space and renal tubules dilation, altered tubular lumen and thyroidization of tubules, , hypertrophy of nucleus, cellular necrosis and atrophy		(Qureshi et al., 2016)
24	Labeo rohita			Rohu		Imidacloprid	Heart, liver, kidney		No histopathological changes observed in heart, Liver : Wrinkling of hepatocytes membrane, hepatocyte degeneration and necrosis, dilation of blood sinusoid, nuclear degeneration, hepatic nuclei' pyknosis, Kidneys : wide Bowman's space, cell necrosis and inflammation, enlargement of renal tubular lumen		(Qadir et al., 2016)
25	Carnivorous species (Chitala chitala, Channa striata, Clupisoma gaura, Wallago attu, Rita rita, Sperata seenghala,), Herbivorous species (Catla catla, Cirrhinus mrigala, Labeo rohita, Cyprinus carpio, Cirrhinus reba, Labeo dyocheilus)			Indian featherback or Indian knifefish, Chitol, Striped Snakehead, River Catfish, Wallago, Rita, Giant River-Catfish, Herbivorous species Major Carp, Mrigal carp, Rohu, Common Carp, Reba Carp, Brahmaputra Labeo, Kali		Organochlorine Pesticides (OCPs) and Polychlorinated Biphenyls (PCBs)	Muscles		OCPs and PCBs detected in edible fish species with highest concentrations recorded in carnivorous species, Health risk to consumers		(Robinson et al., 2016)
26	Cyprinus carpio, Tor putitora, Glyptothorax punjabensis, Orienus plagiostomus			Common carp, Mahseer, Sisorid catfish, Sattar snow trout		DDT (Dicholorodiphenyltricholoroethane), HCHs (Hexachlorocyclohexane)	Muscles, gills, stomach, liver tissues		Bioaccumulation of HCHs and DDT in fish especially in G. punjabensis and C. carpio, DDT intake with life time consumption of mentioned fish species pose cancer risk to local people.		(Aamir et al., 2016)
										Table continued on next page.................
S. No	Fish			Common name		Name of pesticide		Sample		Changes noticed	Reference
27	Ctenopharyngodon idella			Grass Carp		Atrazine (Herbicide)		Blood		Decrease in synthesis and activity of enzymes serum glutamic-pyruvic transaminase (SGPT), creatinine phosphokinases (CPK), lactate dehydrogenase (LDH), and alkaline phosphatase, resulting in enzyme accumulation in cells due to reduced permeability for mentioned enzymes.	(Khan et al., 2016)
28	Oreochromis mossambicus			Mozambique tilapia		Organophosphates (Chlorpyrifos, malathion), Pyrethroids (Cypermethrin, λ-Cyhalothrin), Insecticide		Blood		Increase in MN (Micronucleus) frequencies; Carcinogenic effect)	(Naqvi et al., 2016)
29	Oreochromis mossambicus			Mozambique tilapia		Organophosphates (Chlorpyrifos, malathion), Pyrethroids (Cypermethrin, λ-Cyhalothrin), Herbicide (Buctril)		Fish tissue		Decrease in protein content and metabolic dysfunction in investigated fishes	(Naqvi et al., 2017)
30	Labeo rohita			Rohu		Chlorpyrifos		Blood		Decline in total RBCs count, Hb, packed cell volume, Increase in total leucocyte count, Increase in MN (Micronucleus) induction	(Ismail et al., 2018)
31	Labeo rohita			Rohu		Endosulfan		Blood		DNA damage, genotoxic effects	(Ullah et al., 2017)
32	Channa marulius, Channa punctatus, Labeo boga			Great Snakehead, Spotted Snakehead, Boga, Boga Labeo		Organochlorine pesticides (DDT, endosulfan, andrin)		Fish tissue		Bioaccumulation of pesticide residues in fish tissues, Carcinogenic risk	(Riaz et al., 2018)
33	Cyprinus carpio			Common Carp		Fipronil		Blood		Convulsions, dizziness, fainting, Increased movement of operculum, jerking, body curvature, Breathing difficulties, Decrease in RBCs, Hb, Hct and albumin, Increase in leukocyte count, mean corpuscle volume, neutrophils, monocytes, lymphocytes, urea, creatinine, cholesterol, triglycerides, glucose, nuclear and cellular abnormalities.	(Ghaffar et al., 2018)
34	Labeo rohita			Rohu		Diafenthiuron		Blood		Increase in WBCs, RBCs, lymphocyte, Hb, Hct, MCV, Decrease in platelets count, plateletcrit, platelet distribution width, Disturbed concentration of total proteins, cholesterol, triglycerides, albumin, globulin, AST, calcium, potassium, and cadmium	(Riaz-ul-Haq et al., 2018)
										Table continued on next page.................
S. No	Fish			Common name		Name of pesticide		Sample		Changes noticed	Reference
35	Labeo rohita			Rohu		Cypermethrin		Blood, gills, liver, intestine		Blood: Rise in WBCs, platelets, blood glucose level, Decline in RBCs count, Hct, Hb, MCV, MCH, MCHC, Gills: lamellar disorder, cartilaginous core disruption, epithelial lifting, blood mobbing, fusion, twisting and shortening of secondary lamellae and degeneration and atrophy, Intestines: hemorrhages, intestinal necrosis, goblet cells’ overproduction in villi, disintegration, shortening and fusion of villi, Liver: cell membrane dissolution, pyknosis, hyperplasia, congestion of blood, cellular necrosis, and vacuolization	(Khan et al., 2018)
36	Oreochromis niloticus			Nile Tilapia		Mixture of endosulfan and chlorpyrifos		Blood		Genotoxic threats, DNA damage	(Ambreen and Javed, 2018)
37	Labeo rohita			Rohu		Mixture of endosulfan and chlorpyrifos		Liver, gills, kidneys, brain, heart, muscles		Antioxidant enzymes i.e. Peroxidase, Superoxide dismutase, and Glutathione S-transferase activity increase in all investigated tissues, while catalase activity increased in liver, gills and kidneys, and decreased in brain, heart, and muscles	(Naz et al., 2019)
38	Catla catla,			Major Carp		Profenofos		Muscles		Rapid increase in moisture content, decrease in proteins, fats, and carbohydrate content	(Ghazala et al., 2019)
	Labeo rohita,			Rohu
	Cirrhinus mrigala			Indian Carp
39	Orechromus mossambicus			Mozambique tilapia		Chlorpyrifos		Blood		Biochemical parameters: Decrease in RBCs, Hb, and HCT, Anemic condition, Increase in WBCs and platelets count, Serological parameters: Increase in blood glucose level, cholesterol, cortisol, Decrease in total protein content, triglycerides	(Ghayyur et al., 2019)
40	Ctenopharyngodon idella			Grass Carp		Mixture of endosulfan+chlorpyrifos and endosulfan+bifenthrin		Live fish (behavior) Fish tissues		Behavior: abnormal behavior, fish try to escape, come to surface, gulp air, increased operculum movements, erratic movement, fast swimming, hyperactivity, Catalase activity and Total Protein Content decreases in liver, kidney, brain, heart, muscle, and gills	(Usman et al., 2020)
41	Labeo rohita			Rohu		Thiamethoxam		Live fish (behavior), blood		Behavior: bouncing movement, rapid operculum movement, erratic swimming, mucus secretion, fin tremors and darkening, isolated swimming on just one side, surface breathing, Hemato-biochemical parameters: Decrease in total RBCs count, Hb, packed cell volume, Increase in WBCs and neutrophils count, Morphological changes: stomatocytes, leptocytes, tear-shaped and dividing RBCs, Nuclear changes: micronuclei, RBCs with condensed nuclei and/or without nucleus, nuclear remnant in RBCs, Histopathology of gills: secondary lamellar atrophy, pyknosis of epithelial pillar cells in lamellae, lamellar degeneration, aneurysm, curling and congestion.	(Ghaffar et al., 2020)
										Table continued on next page.................
S. No	Fish			Common name		Name of pesticide		Sample		Changes noticed	Reference
42	Labeo rohita				Rohu	Thiamethoxam		Blood, Liver, Kidneys		Increased concentration of urea, creatinine, lipid peroxidation product, liver and cardiac function tests, blood DNA content, DNA damage in kidneys, liver, and blood cells, Liver: congestion, bile duct degeneration and disruption, condensation, abnormal sinusoids, eccentric nuclei fragmentation in nucleus, fatty infiltration, Kidneys: necrosis, congestion, edema, detachment of tubular epithelium, degeneration of renal tubules and glomerulus	(Hussain et al., 2020)
43	Oncorhynchus mykiss				Rainbow Trout	Chlorpyrifos		Blood, liver, gills		Blood: decline in RBCs, Hb, Hct, and Acetylcholinesterase activity, Rise in WBCs, Increased activity of antioxidant enzymes, Liver: hepatic degeneration, hyperaemia, abnormal sinusoids, dilation of central veins, Gills: fusion and curling of secondary lamella, cellular degeneration, necrosis, edema, hyperplasia, sloughing, narrowed water channels	(Ali et al., 2020; Yaseen et al., 2016)
44	Oreochromis niloticus				Nile Tilapia	Malathion		Live fish, Blood		Enhanced concentrations of urea and creatinine, kidney damage, serological alterations, Behavior: hyper-excitability, erratic movements, active swimming, Morphological changes: bulging of the eyes, over secretion of mucus, hemorrhage of eyes and body, tail rotting, scale erosion, and mortality	(Zulfiqar, 2020)
45	Oreochromis niloticus				Nile Tilapia	Malathion, Chlorpyrifos, λ-Cyhalothrin		Brain, gills, muscles, kidneys, liver, blood		Decrease in total protein content	(Amin et al., 2021)
46	Labeo rohita				Rohu	Fipronil		Liv e fish, blood, visceral organs		Behavior: loss of coordination, increased operculum	(Ghaffar et al., 2021)
										movement, fin tremors, Blood: RBCs, monocytes, lymphocytes decreased, WBCs, neutrophils increased, nuclear abnormalities in RBCs, Histopathology: severe lesions in gills and liver, Increased levels of ALP, AST, ALT and LDH
47	Cirrhinus mrigala		Mrigal carp			Chlorfenapyr, Dimethoate, Acetamiprid		Tissues, blood		Decrease in RBCs, Hb, PCV, MCHC, T3 and T4, Increase in WBCs and Platelet count, TSH, corticol, ALP, AST, ALT. LDH levels, Histopathological alterations in gills and liver	(Ghayyur et al., 2021)
48	Labeo rohita		Rohu			Pyriproxifen (PPF)		Visceral organs, blood		Decline in RBCs, HCT, Hb, Increase in WBCs, neutrophils, and biomarker values of liver, kidneys and heart, Liver: necrosis, inflammatory exudate, edema, Kidneys: tubular necrosis, widening of Bowman's space, edema, Brain: micro-gliosis, degeneration, hemorrhages, neural pyknosis, Heart: edema, neutrophilic myocarditis, cardiac myofibers disruption	(Naseem et al., 2022)
										Table continued on next page.................
S. No	Fish		Common name			Name of pesticide		Sample		Changes noticed	Reference
49	Labeo rohita		Rohu			Pyriproxifen (PPF)		Visceral organs, blood		Nuclear and morphological alterations in RBCs, Increase in miconucleus, pear-shaped RBCs, nuclear remnants, RBCs with a blebbed nucleus, and spherocytes, Increased DNA damage, Rise in quantity of oxidative stress biomarkers, Decline in antioxidant enzymes' concentration	(Li et al., 2022)
50	Aristichthys nobilis		Bighead Carp			Acetochlor		Visceral organs, blood		Isolated cells of kidneys, brain, gills, and liver showed increased DNA damage and low cellular proteins, Increased morphological and nuclear changes, Rise in quantity of biomarkers of oxidative stress	(Mahmood et al., 2022)
51	Aristichthys nobilis		Bighead Carp			Triclosan		Blood, Visceral organs		Behavior: jerking movement, erratic and irregular swimming, mucus secretion from the mouth, Decline in RBCs, Hb, and Hct values, Rise in WBCs, neutrophils, AST, ALT, urea, creatinine, and cardiac biomarkers, increased DNA damage, nuclear and morphological variations in RBCs, Gills: lamellar uplifting and disorganization, twisting of secondary lamellae, and epithelial cell necrosis in lamellae, Liver: congestion, necrosis, fatty infilteration, Kidneys: increased urinary spaces, tubular necrosis, Brain: necrosis, atrophy of neurons,	(Akram et al. 2022)
52	Hypophthalmichthys nobilis	Bighead Carp				Pendimethalin		Visceral organs, blood		Behavior: overproduction of mucus, loss of equilibrium, erratic and irregular swimming, rapid operculum movement, increase in surface breathing, air gulping, and fin tremors, Liver: congestion, necrosis of hepatocytes, and atrophy of hepatocytes, Gills: lamellar atrophy and fusion, congestion, epithelial cell necrosis of primary and secondary lamellae, secondary lamellar uplifting, telogenesis, Kidneys: degeneration of renal tubules, ceroid, atrophy of glomerulus, necrosis of renal tubules, Increased DNA damage , Increased morphological and nuclear changes, Rise in quantity of biomarkers of oxidative stress	(Wang et al., 2022)
53	Ctenopharyngodon idella	Grass Carp				Lambda-cyhalothrin		Blood, gills, muscles, brain, liver		Severe histopathological lesions in investigated organs, alterations in serum biochemistry, disturbed glucose, total protein, triglycerides, and amylase levels	(Niaz et al., 2022)

 

Other insecticides and pesticide

Profenofos (LC50): Profenofos are new insecticide and neuro toxic molecules. Widely used in in India, Pakistan, Bangladesh for agriculture purpose. The pesticide is highly risk for aquatic organism like fish (Lundebye et al., 1997).

λ-cyhalothrin: The λ-cyhalothrin used for Agriculture purposes such vegitable production and cotton cultivation. It observed in running water in irrigation canal.the toxicity clearly indicates acute and sub-acute toxicity test, toxicity of pesticide with species temperature, and size of the fish (Bibi et al., 2014).

Dichlorodiphenyltrichloroethane (DDT): The Dichlorodiphenyltrichloroethane is used pesticide and kill the Aquatic organism (vertebrates and invertebrates). Causes death in fish hatchery kill fry and effect on fish fecundity (Hopkins et al., 1969).

Pyriproxyfen: Pyriproxyfen is most sensitive pesticide, high concentration detected in Daphnia magna, effected reproductive organ such as, Xiphophorus maculatus, Eurytemora affinis, Leander tenuicornis, Danio rerio (zebrafish) and Capitella sp. (polychaete). highly detected from river water and effected 25.82% species (Moura and Souza-Santos, 2020).

Pendimethalin: Thy known as (N-(1-ethylpropyl)-2, 6-dinitro-3,4-xylidine) belong to dinitroaniline herbicide and commonly used in terrestrial system (Danion et al., 2014). High concentratin detected in river (840 ng/L) at France and effected biotic and abiotic components of ecosystem (Dupuy et al., 2019).

Triclosan: The 5-chloro-2-(2,4-dichlorophenoxy)-phenol commonly used in personal care products such as dental care products, deodorants and textiles (Liang et al., 2013). TCS higher concentration recorded in sediments then water. It has androgenic effects potential, altering swimming of (Oryzias latipes), reproductively effects of (Closterium ehrenbergii) and genotoxic effects on C. ehrenbergii has reported (Liang et al., 2013).

Acetochlor: Acetochlor is well known herbicide, effect the aquatic species including Bighead carp, there are different Scientific names base on divergent branchial row abdominal keel and Length, such as Aristichthys nobilis, Cephalus hypothalamus, Leuciscus nobilis, Hypophthalmichthys mandschuricus, and Hypophthalmichthys simony. Hypophthalmichthys nobilis (Ghaffar et al., 2017).

Hexachlorocyclohexane (HCH): Hexachlorocyclohexane is harmful pesticide mostly used in Pakistan. It cause serious health impact including reproductive, neurological, hematological and immunological disorder on animals (Kalyoncu et al., 2009).

Atrazine: Atrazine (2–18 chloro-4-ethylamino-6-isopropylamino-s-triazine) is toxic herbicide. Effect erythrocyte of Lithobates catesbeianus, Dendrophryniscus minutus, Rhinella, schneideri Anaxyrus americanus, Xenopus laevis, Lithobates pipiens (Khan et al., 2012).

Conclusions and Recommendations

This article concludes that increased and unplanned of pesticides renders great threat to environment, aquatic life, and humans. Pesticides causes different behavioral, hematological, histopathological, genotoxic, endocrine alterations in fish body, which is tolerated by fish only up to a certain level beyond which mortality of fish occurs, resulting in economic losses. On the other hand, these affected fishes cause different hematological and endocrine disorders and some even poses carcinogenic risk to humans when consumed. Regulations and awareness among masses should be developed for responsible utilization of pesticides and to control run-off of these pesticides to aquatic bodies. Furthermore, those fishes should be stocked in water bodies which are least vulnerable to pesticides and chemicals. Further research is needed to study the effects of newly introduced pesticides and to develop biodegradable pesticides that are environment friendly and have least toxicity to non-target organisms.

Acknowledgments

The authors are thankful to the supporting staff of the Department of Zoology and College of Veterinary Science and Animal Husbandry (CVS & AH), Abdul Wali Khan University Mardan,( KP) for their assistance.

Novelty Statement

The research and experimental work on the subject title recommends responsible utilization of pesticides and discourage unnecessary use of pesticides in aquatic environment.

Author’s Contribution

Yaseen and Asad Ullah: Designed and prepared the manuscript.

Imad Khan and Maryam Begum: Supervised the write up.

Sumbal Bibi and Umber: Helped in data collection.

Namra and Abbas Khan: Technically assisted at every step.

Shumaila Gul and Raheela Taj: Proof reading.

Conflict of interest

The authors declared no potential conflicts of interest with respect to research, authorship, and/or publication with the work submitted.

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