Does short-term exposure to a Neonicotinoid insecticide trigger Biochemical and Physiological responses in Juvenile catfish?

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Nosakhare O. Erhunmwunse, Timeyin J. Pajiah, Ogbodo Victory, Endurance Ewere This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2193834/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Imidacloprid is among the most widely used insecticides and may contaminate surface waters, yet there is limited information available on their effects on fish. In the present study, juvenile catfish ( Clarias gariepinus ) were exposed to 100, 130, 160, and 190 mg/L of imidacloprid for 96 hours, and the effects on mortality, behavior, hepatic and endocrine functions, oxidative stress, and tumor and inflammatory responses were investigated. The 96 hours LC 50 was 166.60 (143.38-193.59 95% confidence intervals) mg/L with mortality recorded at concentrations ≥ 130 mg/L. Exposure to imidacloprid induced behavioral alterations and clinical symptoms including gulping, hypoactivity, abnormal surface distribution, loss of buoyancy, and excessive mucus secretion. Furthermore, amylase, lipase, conjugate bilirubin, carbohydrate antigen 19 − 9 (CA 19 − 9), thyroid stimulating hormone (TSH), and glutathione peroxidase (GPx) activity were significantly increased, while significant reductions were observed in triiodothyronine (T3) and superoxide dismutase (SOD) activity. Non-significant changes were observed in aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin, 17β- estradiol (E2), thyroxine (T4), C-reactive protein (CRP), glutathione S-transferases (GST), total antioxidant capacity (TAC) and catalase (CAT) activity. Our results demonstrate that imidacloprid causes behavioral, hepatic, and endocrine toxicity in catfish as well as oxidative stress and tumor marker elevation. Our findings show that short-term exposure to imidacloprid triggers biochemical and physiological responses in juvenile catfish. Toxicology Behavioral Ecology Animal Physiology behavioral markers imidacloprid oxidative stress thyroid hormones reproductive hormones hepatic markers tumor marker inflammatory marker Figures Figure 1 Introduction Pollution has taken the headlines in recent stories across the globe as more contaminants with varying magnitudes of toxicity are being introduced into aquatic and terrestrial environments. Man's attempt to improve the quality of life subjects the ecological system to enormous stress and deteriorating modifications. 1 These activities affect the health and survival of individual organisms, as well as their performance, and alter the function and structure of natural ecosystems, especially the aquatic ecosystem. 2 Commercial, industrial, and agricultural chemicals can cause several harmful effects on aquatic organisms when they enter the aquatic environment. Fish accumulate these chemicals, including pesticides, directly from the polluted water and indirectly from the food chain. 3,4 Over a thousand varieties of pesticides are used in the world today, with different properties and exploring diverse modes of action, with more than 80% of farmers relying on pesticides to secure their produce. 5 Some pesticides like DDT and lindane have been proven to be very persistent in soil and water for many years after application. 6 Most pesticides deviate from their main objective of the action, causing a variety of toxic effects in non-target species. 7 As pesticides ultimately flow into the surface water, it is expected that aquatic environments are more affected than terrestrial environments. 8,9 Aquatic ecosystems are threatened by the pollution of these pesticides, thus the development and usage of pesticides with low ecological impact and toxicity to non-target organisms are pertinent for the preservation of these ecosystems and biodiversity. 10 Great efforts have been made over the last three decades to produce pesticides that are specific in action, highly degradable, and have selective qualities that work on certain chemical and physiological aspects of insects, reducing the risk of non-target species. This tremendous advance in knowledge paved the way for a more effective group of insecticides, the neonicotinoids, of which imidacloprid is a widely used example. Neonicotinoids are a class of nitroguanidine systemic insecticides often used to treat soil and seeds before planting crops to prevent pest infestations. 11 Owing to their effectiveness against a wide range of insects and the breadth of their uses, as well as their functioning at low dosages with relatively low volatility and high solubility in water. 12,13,14 The idea that neonicotinoids function as nicotinic acetylcholine receptor (nAChR) agonists, which are thought to have little impact on vertebrate species and limited cross-resistance with other pesticides, is the basis for their appeal. Also, neonicotinoids have systemic activity, which allows them to spread more widely throughout plants than conventional pesticides to control the targeted pest species. 10 However, neonicotinoids' high water solubility also makes them very mobile and, as a result, easily transferred into aquatic habitats, potentially causing harm to aquatic organisms that are not intended targets. Seven neonicotinoid pesticides are currently in use, including acetamiprid, dinotefuran, clothianidin, nitenpyram, imidacloprid, thiamethoxam, and thiacloprid. Imidacloprid [1-(6-chloro-3-pyridylmethyl)-N-nitro-imidazolidin-2-ylideneamine] is a neonicotinoid synthetic insecticide that forms an agonistic relationship with the postsynaptic nicotinic acetylcholine receptors 15,16 causing impairment of normal nerve function. It was the first neonicotinoid to be introduced in 1991, and it has since become a key product in several pest control operations. 17,18 Products containing imidacloprid dominate the insecticide industry and are authorized for use in more than 120 countries on more than 140 different types of crops. 19 Its continuously increasing popularity not just in the control of household pets’ pests but especially in agricultural use can be attributed to its excellent effectiveness in the control of sucking, soil, and chewing insects. 20 Recently, numerous studies have focused on the toxicity of imidacloprid in the aquatic environment due to the negative effects of neonicotinoids on non-target beneficial invertebrates. Although there is little information available about its toxicity on aquatic vertebrates including fish, several studies have assessed the toxic effects of imidacloprid on several endpoints of different species of fish in various parts of the world. 14,21,22 Imidacloprid has been reported to cause DNA damage and oxidative stress 14 decreased hatching rate of fertilized eggs 22 and histopathological alterations in tissues 23 of different fish species. Nonetheless, no study has conducted a broad range of assessments of behavioral and physiological responses and biomarkers for a fish species. This is necessary to understand the underlying mechanism of action for imidacloprid toxicity in fish and other vertebrates. Considering these knowledge gaps, this study aimed to assess the impacts of imidacloprid on Clarias gariepinus behavior, hepatic and endocrine functions, oxidative stress, and tumor and inflammatory responses. To achieve this objective, catfish were exposed to acute concentrations of imidacloprid for 96 hours. C. gariepinus was chosen for this study due to its wide distribution in tropical freshwater systems, adaptation to laboratory conditions, and stress resistance. 24 Materials And Methods Fish Maintenance. Juvenile Clarias gariepinus (mean length = 10.73 ± 1.34 cm; mean weight = 12.34 ± 3.44 g) were purchased from Farm Project, Faculty of Agriculture, University of Benin, Nigeria and maintained in a 160 L plastic tank for two weeks according to OECD 25 at the Laboratory for Ecotoxicology and Environmental Forensics at the University of Benin, Nigeria. Chemical Exposures. Chemical exposure was conducted through a static renewal system for 96 hours, following OECD guidelines for fish acute toxicity testing 25 . The treatment groups consisted of four concentrations of imidacloprid: 100, 130, 160, and 190 mg/L using a commercially available insecticide, Imi Force® (Jubaili™), containing an imidacloprid concentration of 200 g/L, and a control group. The treatment concentrations were achieved by mixing 10, 13, 16, and 19 mL of the Imi force® insecticide in 20 L water to get the required imidacloprid concentrations of 100, 130, 160, and 190 mg/L respectively. No insecticide or chemical was added to the control group. The concentrations and duration were selected based on previous studies and a range-finding test. Each group was comprised of two replicates (n = 6). Mortalities were counted daily for LC 50 determination. Behavioral Alterations and Mortality. The fish were inspected multiple times daily during the exposure period to record behavioral changes and clinical diagnostic signs through direct observation, according to OECD 25 . The toxicological endpoint of this study was mortality. Mortality was recorded daily when the fish failed to respond to gentle prodding. The dead fish were removed immediately. Sampling. All fish were euthanized after 96 hours of exposure to a lethal dose of benzocaine (0.5 g L − 1 ; Sigma-Aldrich). Samples were selected randomly from each aquarium (n = 2), homogenized, and used for the assays. Biomarker Analysis. The samples were used in the assay of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities according to Reitman and Frankel 26 ; total and conjugate bilirubin levels according to Jendrassik and Grof 27 ; amylase activity according to Bern 28 ; lipase activity according to Bier 29 ; 17β-estradiol (E2) and testosterone levels using an Estradiol EIA Kit (Cayman Chemical, USA) according to the manufacturer’s instructions; glutathione S-transferases (GST) activity according to Habig and Jakoby 30 ; carbohydrate antigen (CA-19-9) level according to Minamide et al . 31 ; C-reactive protein (CRP) level according to Lowry et al. 32 ; total antioxidant capacity (TAC) activity according to Teimouri et al . 33 ; catalase (CAT) activity according to Beer and Sizer 34 ; glutathione peroxidase-1 (GPx) activity according to Hafemann et al. 35 ; superoxide dismutase (SOD) according to McCord and Fridovich 36 ; triiodothyronine (T3), thyroxine (T4) and thyroid stimulating hormone (TSH) using ELISA kit (Millipore-Sigma, USA) according to the manufacturer’s instructions. Statistical Analysis. LC 50 was calculated with probit analysis using Microsoft Excel 2016. Statistical analyses were conducted with GraphPad Prism (version 5.01). All data were analyzed using a single factor one-way analysis of variance (ANOVA), followed by Dunnett’s posthoc test. Statistical significance was determined if P < 0.05. Results And Discussion Fish are ideal sentinels for behavioral and health assays of various stressors and toxic chemical exposure due to their constant direct contact with the aquatic environment, where chemical exposure occurs over the entire body surface. Imidacloprid is a promising and rapidly expanding pesticide, known for its selective toxicity to insects. Its detection in food and water prompts the verification of its possible negative effects on non-target organisms. To identify its effect on Clarias gariepinus , various biomarkers were evaluated along with mortality and behavioral changes. This present study contributes to the ongoing efforts to unravel the adverse effects and mode of action of imidacloprid and other neonicotinoid pesticides on non-target species. This study is the first to report behavioral alterations and endocrine disruption in Clarias gariepinus after imidacloprid exposure, as well as the effects of imidacloprid on Ca 19-9 in fish. The results of this study show that exposure to different imidacloprid concentrations adversely impacted C. gariepinus . Imidacloprid LC 50 , Mortalities, Fish Behavior, and Clinical Symptoms. The water temperature and pH in the aquariums throughout the toxicity testing were 26.64 ± 0.09 ℃ and 6.74 ± 0.05 respectively. The survival rate of fish in the control was 100%, indicating that the toxicity data were acceptable. The mortality rate was time- and concentration-dependent, with % mortality peaking at 66.67% after 96 hours of exposure. Mortality was observed at concentrations ≥130 mg/L. (Table 1). The 96-h LC 50 of imidacloprid was 166.60 mg/L (95% confidence intervals: 143.38-193.59 mg/L). This quantity is substantially higher than the environmental values already measured and is unlikely to occur in aquatic systems. The value obtained in this study is considerably higher than the previously reported 96-h LC 50 (10.16 µg/L) for C. gariepinus . 21 However, similar values have been reported for imidacloprid in other fish species, including Danio rerio (143.7 mg/L) 37 and Misgurnus anguillicaudatus (145.8 mg/L) 38 . Table 1. Cumulative mortality of Clarias gariepinus exposed to various concentrations of imidacloprid Imidacloprid concentration (mg/L) Number of exposed fish Cumulative mortality of fish per time interval (h) Mortality % 24 48 72 96 0 12 0 0 0 0 0 100 12 0 0 0 0 0 130 12 0 0 1 3 25 160 12 1 1 3 5 42 190 12 3 6 7 8 67 Fish behavior varied with concentration and time of exposure. All exposed fish showed noticeable distress symptoms, represented by under-reactivity to stimulus, hypoventilation, gulping, abnormal surface distribution, and poor swimming movement (Table 2). Loss of buoyancy was also observed in fish, a few hours before mortality. Fish exposed to ≥ 130 mg/L imidacloprid were severely hypoventilated, exhibiting almost no opercular movement, while fish in ≥ 160 mg/L constantly gasped for air at the water's surface. This could be a result of imidacloprid affecting fish gills directly, which may affect fish respiration. 39 Although fish activity was largely reduced in all exposed fish compared to the control, fish activity in the 100 mg/L groups showed improvement on the final day of observation. Furthermore, mucus secretion in all exposed fish peaked after 48 hours of exposure but declined after 72 hours. These observations of fish activity and mucus secretion suggest that catfish may recover from acute behavioral effects caused by imidacloprid after a prolonged time of exposure. Literature on the effects of imidacloprid on fish behavior is scarce, however, imidacloprid has been reported to cause similar behavioral alterations by Qadir et al . 40 in Labeo rohita exposed to 120 mg/L imidacloprid for 8 days. Qadir et al . 40 also observed that as exposure time increased, fish tended to recover from the disturbing condition and the occurrence of abnormal behavior declined, similar to our findings. The behavioral changes observed in our study could be attributed to the effect of imidacloprid on the fish nervous system via impairment of brain activity and acetylcholinesterase level alteration, resulting in the impediment of many physiological functions. 41,42 Table 2. Behavioral abnormalities of C. gariepinus due to the exposure to imidacloprid within the 96-h observation period Concentration (mg/L) Hypo activity Abnormal surface distribution Under-reactivity to stimuli Hypoventilation Gulping Mucus secretion 24h 0 - - - - - - 100 +++ +++ ++ +++ ++ ++ 130 +++ +++ +++ +++ ++ ++ 160 +++ +++ +++ +++ +++ +++ 190 +++ +++ +++ +++ +++ +++ 48h 0 - - - - - - 100 +++ +++ ++ +++ ++ ++ 130 +++ +++ ++ +++ +++ ++ 160 +++ +++ +++ +++ +++ +++ 190 +++ +++ +++ +++ +++ +++ 72h 0 - - - - - - 100 +++ +++ ++ +++ ++ + 130 +++ +++ +++ +++ +++ + 160 +++ +++ +++ +++ +++ ++ 190 +++ +++ +++ +++ +++ ++ 96h 0 - - - - - - 100 ++ ++ ++ +++ ++ - 130 +++ +++ +++ +++ +++ - 160 +++ +++ +++ +++ +++ - 190 +++ +++ +++ +++ +++ - '-' indicates no behavioral abnormality; '+' indicates the number of individuals showing behavioral abnormalities; '+'= Mild (around 30 to 40% of individuals showing behavioral abnormalities); '++' = Moderate (around 45 to 65% of individuals showing the behavioral abnormalities); ‘+++’ = Severe (≥ 70% of individuals showing the behavioral abnormalities). Hepatic Biomarkers. Concentrations of ALT and AST were increased in fish exposed to imidacloprid, however, these changes were not significantly different from the control. ALT levels increased with increasing concentrations of imidacloprid until 160 mg/L peaking at a 176% difference from the control. AST levels increased considerably in fish exposed to imidacloprid, with a difference of more than 17 folds observed at 130 mg/L (Table 3). The increased enzymatic activities could result from elevated enzyme production to counter the harm caused by the insecticide or a disturbance in the cell membrane integrity, preventing the enzymes from moving freely. 43,44,45 Increases in ALT and AST levels have previously been reported in C. gariepinus and other fish species exposed to neonicotinoids. 40,46,47 Desai and Parikh 46 studied the biochemical effects of imidacloprid on Labeo rohita and Oreochromis mossambicus . Fish exposure to imidacloprid for 21 days elevated AST and ALT levels in several tissues of Oreochromis mossambicus and Labeo rohita . 46 Increased activities of ALT and AST could be due to amplified transamination processes that occur as a result of amino acid input into the tricarboxylic acid cycle to manage energy crises during toxicant-based stress. 48,49 The likelihood of the elevation of the enzymes due to an energy crisis is supported by our observations on amylase and lipase activity in exposed groups. Amylase activity in catfish increased with increasing concentrations of imidacloprid. A significant increase in amylase activity was observed in catfish exposed to 190 mg/L imidacloprid. Similarly, a concentration-dependent increase was observed in lipase activity. A significant difference from the control was observed in fish exposed to ≥ 130 mg/L with the confidence intervals increasing with each observed concentration of imidacloprid (Table 3). Amylase and lipase are important enzymes that are directly related to carbohydrate and lipid metabolism. Amylase is secreted by the exocrine region of the pancreas. The elevated activity could be the result of amylase secretory cell damage or pancreatitis. 45 Alteration in serum amylase indicates an abnormality of the digestive process. 50 In biological systems, lipase is crucial for maintaining the integrity of membranes and cell signaling. 51 Increased lipase activity could be due to tissue damage, the development of a stress situation, or the effective use of lipid molecules for digestion to match the high energy demand during the imidacloprid-induced stress condition. 52 Literature available on the effects of imidacloprid on lipase and amylase activities in fish is limited, however, increases in their activities in several fish species due to other pesticides have been reported. 50,51,52,53 Our study indicates that imidacloprid can impact fish metabolic activities, forcing increased energy production through carbohydrate and lipid metabolism. No significant difference was observed between total bilirubin levels in control and catfish exposed to imidacloprid. The total bilirubin levels in exposed catfish were generally higher than the control, with a drop below the control level observed only in the 190 mg/L groups. In contrast, a significant difference from the control was observed in conjugate bilirubin levels of the fish exposed to 130 mg/L. However, similar to the patterns observed in total bilirubin levels, conjugate bilirubin levels of catfish exposed to imidacloprid were generally higher than that of the control, with a drop below control levels observed only in the 190 mg/L groups (Table 3). Bilirubin is one of the main bile pigments found in the fish circulatory system and is derived from hemoglobin disruption. 54 The increased bilirubin production could result from imidacloprid inducing increased red blood cell breakdown or reduced hepatic uptake of bilirubin due to the decreased transport protein-albumin level 55 , severe damage to hepatocytes, and hindrance of the bile duct. 56 Increase in total and conjugate bilirubin levels have also been linked to liver and biliary tract disease. 57 Experimental studies have shown that exposure of C. gariepinus and other fish species to neonicotinoids, including imidacloprid, causes an increase in bilirubin levels 40,58 which among others has been attributed to acute hepatic necrosis. 58 Table 3. Effects on hepatic functions of C. gariepinus after 96 hours of exposure to imidacloprid Imidacloprid (mg/L) ALT (U/l) AST (U/l) Amylase (U/l) Lipase (U/l) Total bilirubin (µmol/l) Conjugate bilirubin (µmol/l) 0 3.38 ± 1.24 4.45 ± 0.15 3.70 ± 1.60 0.15 ± 0.03 0.31 ± 0.04 0.06 ± 0.02 100 7.51 ± 3.22 11.75 ± 1.05 7.95 ± 0.55 0.44 ± 0.15 0.37 ± 0.04 0.14 ± 0.01 130 8.58 ± 2.15 78.22 ± 18.22 10.05 ± 4.75 0.55 ± 0.03* 0.58 ± 0.31 0.28 ± 0.07* 160 9.35 ± 2.92 67.50 ± 20.40 17.45 ± 4.75 0.70 ± 0.03** 0.37 ± 0.14 0.18 ± 0.01 190 6.43 ± 0.00 66.45 ± 21.45 23.80 ± 1.60* 1.13 ± 0.01*** 0.12 ± 0.01 0.06 ± 0.01 P value 0.4537 0.0541 0.0340 0.0013 0.4383 0.0189 The values are mean ± standard error of two fish replicates. *, ** and *** indicate significant difference between exposure and control group at p < 0.05, 0.01 and 0.001 respectively. Endocrine Disrupting Biomarkers. Catfish 17β-estradiol (E2) levels increased with increasing concentrations of imidacloprid, having an 85% increase from control levels in the group exposed to 190 mg/L. However, these increments were not significantly different from the control. Testosterone levels in catfish fluctuated minimally with increasing concentrations of imidacloprid, with a maximum 5% difference from the levels observed in the control fish. These changes were not significantly different from the control as well (Table 4). The primary internal regulators of gonadal differentiation and its activities are sex steroid hormones. 59 They influence how the brain is organized during the formative stages and sex-dependently coordinate a range of behaviors. 60 They also have an impact on the neurological, skeletal, and cardiovascular systems, as well as stress and immunological responses. 61,62 E2 is a major sex hormone that controls female fish maturity, and high E2 levels are a reliable sign of endocrine disturbance. 63 Elevated E2 concentrations are especially worrisome because they can upset the endocrine system's delicate balance, impairing reproductive function and resulting in lower fecundity and fertility, repressed sexual behavior, and altered sexual differentiation. 63 Fish immune systems and numerous leukocyte functions are regulated by estrogens. 64 Acidophilic granulocytes, which have been linked to tissue damage 65 , migrate from the head kidney as a result of an increase in E2 levels following an inflammatory reaction. 66 This is the first study on the effects of imidacloprid on catfish sex steroid hormones. Experimental studies have shown that Clarias and several fish species' exposure to pesticides and other stressors cause elevations in E2 levels. 63,67,68,69,70 Additional research is necessary to identify whether imidacloprid affects E2 levels directly or indirectly through the conversion of testosterone to E2, slowed sex steroid hormone degradation mechanisms, or at the hypothalamic-pituitary-gonad axis. The findings indicate that additional research is required to fully understand the negative effects of imidacloprid on the reproductive endocrine axis, regardless of the mode of action that led to the elevated E2 reported in this study. Significant differences were observed between T3 levels in the control and catfish exposed to 190 mg/L. T3 and T4 levels were generally reduced with increasing concentrations of imidacloprid. However, no significant difference was observed between T4 levels in control and fish exposed to imidacloprid. In contrast, TSH levels of fish increased on exposure to imidacloprid, with a significant difference observed between the control and 190 mg/L groups (Table 4). Thyroid hormones in fish regulate growth, development, metabolism, skin pigmentation, behavior, metamorphosis, osmoregulation, and lipid and protein metabolism, often in association with growth hormone and cortisol. 71,72,73,74 Production of T3 is largely by peripheral enzymatic monodeiodination of T4 majorly in the liver and other tissues. 75 Typically, a decrease in T4 synthesis and secretion is what causes T3 levels to decrease. 71 In the present study, a decrease in the level of thyroid hormones might be indicative of metabolic energy redirecting away from the anabolic process, which is essential to carry through life, thereby triggering thyroid endocrine disruption. 76 Thyroid hormone levels are regulated by the negative feedback that thyroid hormones have on their upstream regulators. Reductions in T3 or T4 enhance the production of TSH by the pituitary gland, hence our findings could be a reflection of this mechanism. Ghayyur et al . 77 reported similar findings in their 30-day exposure of Cirrhinus mrigala to the neonicotinoid acetamiprid. They observed significant increases in thyroid hormones and a significant reduction in TSH levels in their experiment after fish exposure to acetamiprid, chlorfenapyr, and dimethoate. Saha et al . 78 observed a reduction in thyroid hormone levels in Clarias batrachus after chronic exposure to diazinon and suggested these reductions could be due to pesticide effects on thyroid hormone signaling and metabolism in the hypothalamic-pituitary-thyroid axis. 78,79 Our study highlights the disruptive endocrine properties of imidacloprid, confirming its potential to adversely affect normal ecological functions and the survival of fish. Table 4. Effects on endocrine functions of C. gariepinus after 96 hours of exposure to imidacloprid Imidacloprid (mg/L) 17β-estradiol (ng/ml) Testosterone (ng/ml) T3 (ng/ml) T4 (µg/dL) TSH (µlm/mL) 0 68.60 ± 4.00 0.80 ± 0.01 1.05 ± 0.05 7.40 ± 1.30 0.70 ± 0.00 100 106.30 ± 15.95 0.80 ± 0.01 1.00 ± 0.00 6.50 ± 0.80 0.80 ± 0.00 130 108.70 ± 16.20 0.84 ± 0.01 1.05 ± 0.05 5.45 ± 0.25 0.80 ± 0.00 160 111.50 ± 5.65 0.81 ± 0.02 0.90 ± 0.00 5.35 ± 0.75 0.80 ± 0.00 190 126.60 ± 17.40 0.78 ± 0.01 0.85 ± 0.05* 4.35 ± 0.35 0.95 ± 0.05** P value 0.1561 0.0551 0.0448 0.2019 0.0047 The values are mean ± standard error of two fish replicates. * and ** indicate a significant difference between the exposure and control group at p < 0.05 and 0.01 respectively. Oxidative Biomarkers. Significant reductions were observed between SOD activity in control and exposed fish at different concentrations of imidacloprid. GPx activity of fish increased in groups exposed to ≥ 130 mg/L imidacloprid. A significant difference between GPx activity in control and exposed fish was observed in 130 mg/L imidacloprid concentration. A non-significant increase in CAT, TAC, and GST activity of the exposed groups compared to the control group was observed (Table 5). Pesticide exposure has the potential to cause oxidative damage. 80 To deal with oxidative stress and prevent excessive reactive oxygen species (ROS) formation, which can cause oxidative damage to nucleic acids, lipids, and proteins, fish produce a variety of anti-oxidative defenses including GPx, CAT, and SOD. 21,41,81,82 The role of GST is to catalyze the detoxification of harmful compounds via the conjugation of glutathione. 83 The current study demonstrated reduced SOD, and elevated GPx, CAT, GST, and TAC in C. gariepinus exposed to imidacloprid. An increase in CAT, GPx, and TAC activity could be an indication that imidacloprid caused an overproduction of ROS in exposed fish. 14 The reduced SOD activity may be due to the elevated oxygen free radicals in catfish, rendering SOD inactive through oxidation. 84 Additionally, elevated quantities of H 2 O 2 and superoxide anions may be the cause of the rise in CAT activity. 85 The increase in H 2 O 2 levels is supported by Svensson et al . 86 who reported that the sulfhydryl of microsomal GST’s 49th cysteine could specifically combine with H 2 O 2 , thus enhancing GST activity. Nnadi et al . 21 reported significant SOD activity reductions and CAT activity elevations in C. gariepinus exposed to imidacloprid, similar to our findings. However, contrary to our results, they found that imidacloprid induced a significant reduction in GPx activity in exposed catfish. Several studies have reported oxidative stress induced by imidacloprid in various fish species such as Danio rerio 87 , Oncorhynchus mykiss 41 , Oreochromis niloticus 88 , and Prochilodus lineatus 14 . Our study further affirms the potential of imidacloprid to induce oxidative stress in fish. Table 5. Effects on oxidative stress of C. gariepinus after 96 hours of exposure to imidacloprid Imidacloprid (mg/L) SOD (U/ml) GPx (Unit/ml) CAT (mmol/min/ml) TAC (mmol/L) GST (mmol/mi/min) 0 1.81 ± 0.07 11.20 ± 2.80 22.30 ± 3.10 1.59 ± 0.16 0.55 ± 0.05 100 1.44 ± 0.14 11.20 ± 0.00 44.60 ± 12.80 1.92 ± 0.01 1.40 ± 0.40 130 1.26 ± 0.08* 36.45 ± 5.65* 36.95 ± 6.85 1.80 ± 0.04 1.20 ± 0.10 160 1.35 ± 0.08* 28.00 ± 2.80 40.15 ± 4.55 1.81 ± 0.05 3.60 ± 2.60 190 1.58 ± 0.00 33.65 ± 8.45 40.35 ± 3.15 1.81 ± 0.13 3.15 ± 2.05 P value 0.0244 0.0353 0.3369 0.3080 0.5814 The values are mean ± standard error of two fish replicates. * indicates a significant difference between the exposure and control group at p < 0.05. Tumor and Inflammatory Biomarkers . Significant increases ( p = 0.0270) between CA 19-9 levels in control and fish exposed to imidacloprid were observed at imidacloprid concentrations ≥ 160mg/L (Figure 1A). Increases in serum CA 19-9 are helpful in the diagnosis of upper gastrointestinal tract adenocarcinoma and the monitoring of colonic carcinoma, but their greatest sensitivity has been found in the detection of pancreatic adenocarcinoma, according to various studies. 89,90 This is the first study assessing pesticide effects on fish CA 19-9 levels. Literature on the effects of toxicants on CA 19-9 in animal studies is incredibly scarce, however, Omnia et al . 91 observed a significant increase in CA 19-9 levels in rats exposed to 1,2-dimethylhydrazine, similar to our results. Our findings indicate that imidacloprid may pose a high risk of malignant and pancreaticobiliary diseases in fish. However, Kim et al . 92 found CA-19-9 elevation in patients with pulmonary, gynecologic, endocrine, and splenic diseases who did not have a malignant and pancreaticobiliary disease. There was no significant difference ( p = 0.0807) between CRP levels in the control and imidacloprid-exposed groups. Furthermore, the CRP levels fluctuated, and the comparison of these two groups did not yield a consistent pattern over time, having only minor deviations from the control group levels except in the group exposed to 190 mg/L (Figure 1B). As an initial line of defense against environmental contaminants, toxic chemicals cause fish to produce more non-antibody proteins like CRP. 93 CRP is mostly produced in the liver and is responsible for several immunological responses. 94 Our studies reveal that although imidacloprid has a mild effect on CRP levels at concentrations below the 96-h LC 50 , its effects become more noticeable at higher concentrations, where a 32% increase in CRP levels was observed in catfish exposed to 190 mg/L imidacloprid. Currently, there is no comprehensive data available on CRP levels due to imidacloprid and other neonicotinoid exposure in fish with which to compare this data. However, elevated CRP levels have been observed in Clarias species on exposure to various stressors. 95,96 Chatterjee 96 exposed C. gariepinus to endosulfan for 96 hours and observed a steady rise in CRP levels. Furthermore, pesticides, as well as other physiological and chemical stressors, have been reported to increase CRP levels in several fish species. 93,97,98,99 This elevated level of CRP in the acute phase response indicates that fish exposed to imidacloprid may develop inflammatory responses. In conclusion, short-term exposure to imidacloprid, a neonicotinoid insecticide, now largely used worldwide as a replacement for more toxic and persistent pesticides triggered biochemical and physiological responses in juvenile catfish ( Clarias gariepinus ). The present study clearly showed that imidacloprid has the potential to disrupt various physiological and behavioral functions in fish, which could lower the probability of survival. While extensive research has focused on the effects of imidacloprid on fish behavior, transaminases, and oxidative stress, our work highlights the need for further examination of its effects on hepatic and endocrine functions, as well as tumor and inflammatory responses. Whether or not the environmentally relevant concentrations of imidacloprid will cause these same effects in catfish remains yet to be known. Controlled use of imidacloprid-based pesticides, especially close to aquatic ecosystems, is therefore essential. Declarations Ethics approval and consent to participate Ethical approval for this research was given by the Research Ethics committee of the College of Medicine of the University of Benin. CMS/REC/2020/34. Consent for publication Not applicable. Availability of data and material The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request. Conflict of interest The authors declare no conflict of interest. Funding This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. CRediT authorship contribution statement Nosakhare O. Erhunmwunse: Conceived and designed the experiments . Timeyin J. Pajiah, Junior V. Ogodo performed the experiments . Nosakhare O. Erhunmwunse, Timeyin J. Pajiah, Junior V. Ogodo: Analyzed and interpreted the data . Nosakhare O. Erhunmwunse, Timeyin J. Pajiah, Junior V. Ogodo, Endurance Ewere: Materials, analysis tools, and ideas . All authors have read and approved the manuscripts. Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Acknowledgment We appreciate the diligence, and relentless efforts of Mr. Christopher who helped us all through the work. References (1) Lopez-Barea, J. (1995). Biomarkers in ecotoxicology . Archives of toxicology, 17 : 57-79. (2) Yancheva, V., Velcheva, I., Stoyanova, S and Georgieva, E. (2016). Histological biomarkers in fish as a tool in ecological risk assessment and monitoring programs: A review. Applied Ecology and Environment Research, 14 (1): 47-75. (3) Mohamed, A. S. F. (2009). Histopathological studies on Tillapia zilli and Solea vulgaris from Lake Qarun, Egypt. World Journal of Fish and Marine Sciences, 1 : 29-39. (4) Chaudhry, A.S., and Jabeen, F. (2011). Assessing metal, protein and DNA profiles in Labeo rohita from the Indus River in Mianwali, Pakistan . Environmental Monitoring and Assessment , 174 : 665-679. (5) Shahzad, F., Taj, M. K., Abbas, F., Taj, I., Parveen, S., Achakzai, A. M., Azam, S., Hussain, A., Tareen, A. R., Mohammad, G., Samreen, Z., Sazian, B. and Bibi, L. (2019). Pesticides and our environment. International Journal of Biosciences, 14 (4): 487-491. (6) Hadian, Z., Eslamizad, S. and Yazdanpanah, H. (2019). Pesticide Residues Analysis in Iranian Fruits and Vegetables by Gas Chromatography-Mass Spectrometry. Iranian Journal of Pharmaceutical Research 18 : 275. (7) Maton, S.M., Dodo, J.D., Nesla, R.A. and Ali, A.Y. (2016). Environmental Impact of Pesticides Usage on Farmlands in Nigeria. International journal of innovative research and development 5 : 311-317. (8) Pritchard, J. B. (1993). Aquatic toxicology: past, present, and prospects. Environmental Health Perspectives, 100 : 249–257. (9) Al-Badran, A. A., Fujiwara, M. and Mora, M. A. (2019). Effects of insecticides, fipronil and imidacloprid, on the growth, survival, and behavior of brown shrimp Farfantepenaeus aztecus . PLoS ONE, 14 (10):e0223641. (10) Hayasaka, D., Suzuki, K., Nomura, T., Nishiyama, M., Nagai, T., Sánchez-Bayo, F. and Goka, K. (2013). Comparison of acute toxicity of two neonicotinoid insecticides, imidacloprid and clothianidin, to five cladoceran species. Journal of Pesticide Science , 38 (1): 44–47. (11) Morrissey, C. A., Mineau, P., Devries, J. H., Sanchez-Bayo, F., Liess, M., Cavallaro, M. C. and Liber, K. (2015). Neonicotinoid contamination of global surface waters and associated risk to aquatic invertebrates: A review. Environment International, 74 : 291-303. (12) EXTOXNET, (1996). Extension Toxicology Network. Pesticide Information profiles: Imidacloprid. Available at http://extoxnet.orst.edu/pips/imidaclo.htm, accessed on 13 April 2022. (13) Tomlin, C. D. S. (2000). The Pesticide Manual, twelfth ed. British Crop Protection Council, Surrey, United Kingdom. (14) Vieira, C. E. D., Pérez, M. R., Acayaba, R. D., Raimundo, C. C. M. and Martinez, C. B. R. (2018). DNA damage and oxidative stress induced by imidacloprid exposure in different tissues of the Neotropical fish Prochilodus lineatus . Chemosphere, 195 : 125-134. (15) Radwan, M. A. and Mohamed, M. S. (2013). Imidacloprid induced alterations in enzyme activities and energy reserves of the land snail, Helix aspersa. Ecotoxicology and Environmental Safety, 95 : 91-97. (16) Shan, Y., Yan, S., Hong, X., Zha, J. and Qin, J. (2020). Effect of imidacloprid on the behavior, antioxidant system, multixenobiotic resistance, and histopathology of Asian freshwater clams ( Corbicula fluminea ). Aquatic Toxicology, 218 : 105333 (17) Jeschke, P., Nauen, R., Schindler, M. and Elbert, A. (2011). Overview of the status and global strategy for neonicotinoids. Journal of Agriculture and Food Chemistry 59 : 2897-2908. (18) Goulson, D. (2013). Review: an overview of the environmental risks posed by neonicotinoid insecticides. Journal of Applied Ecology 50 : 977-987. (19) Jeschke, P. and Nauen, R. (2008). Neonicotinoids—from zero to hero in insecticide chemistry. Pest Management Science 64 : 1084-1098. (20) Lewis, K. A., Tzilivakis, J., Warner, D. J. and Green, A (2016). An international database for pesticide risk assessments and management. Human and Ecological Risk Assessment: An International Journal , 22 :1050-1064. (21) Nnadi, J.U., Dimelu, I.N., Nwani, S.I., Madu, J.C., Atama, C.I., Attamah, G.N., Okwor, J.I. and Nwani, C.D., 2018. Biometric variations and oxidative stress responses in juvenile Clarias gariepinus exposed to Termex®. African Journal of Aquatic Science , 43 (1), pp.27-34. (22) Islam, M. A., Hossen, M. S., Sumon, K. A. and Rahman, M. M. (2019). Acute toxicity of imidacloprid on the developmental stages of Common Carp Cyprinus carpio . Toxicology and Environmental Health Sciences , 11 (3): 244-251. (23) Qadir, S. and Iqbal, F. (2016). Effect of subleathal concentrtion of imidacloprid on the histology of heart, liver and kidney in Labeo rohita . Pakistan journal of pharmaceutical sciences, 29 :2033-2038. (24) Ogueji, E. O., Nwani, C. D., Iheanacho, S. C., Mbah, C. E., Okeke, O. C. and Yaji, A. (2018). Acute toxicity effects of ibuprofen on behaviour and haematological parameters of African catfish Clarias gariepinus (Burchell, 1822). African Journal of Aquatic Science, 43 (2): 293–303. (25) OECD (2019). Test guideline No. 203: Fish, acute toxicity test. OECD Guidelines for the Testing of Chemicals, Section 2. OECD Publishing, Paris. doi:10.1787/9789264069961-en (26) Reitman, S. and Frankel, S. (1957). A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. American journal of clinical pathology, 28 (1): 56-63. (27) Jendrassik, L. and Grof, P. (1938). Estimation of total serum bilirubin level by spectrophotometrically in serum and plasma. Biochem Zeitschrift 297 : 81-89. (28) Bern, F. P. (1955). Amylase a and b. In Methods of enzymology (Ed. Clockwi and Kalpin). Academic Press New York, USA. pp. 149-158. (29) Bier, M. (1955). [106] Lipases: RCOOR′+ H2O→ RCOOH+ R′ OH. (30) Habig, W.H. and W.B. Jakoby (1981). Assays for differentiation of glutathione-S-transferases. Methods in Enzymology, 77 : 398-405. (31) Minamide, M., Hosoi, I. and Yanagi, S. (2000). CA19.9-producing testicular tumor: a case report, Hinyokika Kiyo , 46 (1): 45-47. (32) Lowry, O. H., Rosenbrough, N. J., Farr, A. L. and Randall, R. J. (1951). Protein measurement with the folin phenol reagent. Journal of Biological Chemistry, 193 : 265-275. (33) Teimouri, M., Yeganeh, S. Mianji, G.R., Najafi, M. and Mahjoub, S. (2019). The effect of Spirulina platensis meal on antioxidant gene expression, total antioxidant capacity, and lipid peroxidization of rainbow trout ( Oncorhynchus mykiss ). Fish Physiology and Biochemistry , 45 (3): 977-986. (34) Beers, R. F. and Sizer, I. W. (1952). A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. Journal of Biological Chemistry, 195 (1): 133-140. (35) Hafeman, D.G., Sunde, R.A. and Hoekstra, W.G. (1974). Effect of dietary selenium on erythrocyte and liver glutathione peroxidase in the rat. The Journal of nutrition , 104 (5): 580-587. (36) McCord, J. M. and Fridovich, I. (1969). Superoxide dismutase: an enzymic function for erythrocuprein (hemocuprein). Journal of Biological Chemistry, 244 (22): 6049-6055. (37) Wang, Y., Yang, G., Dai, D., Xu, Z., Cai, L., Wang, Q. and Yu, Y. (2017). Individual and mixture effects of five agricultural pesticides on zebrafish ( Danio rerio ) larvae. Environmental Science and Pollution Research , 24 (5): 4528-4536. (38) Xia, X., Xia, X., Huo, W., Dong, H., Zhang, L. and Chang, Z. (2016). Toxic effects of imidacloprid on adult loach ( Misgurnus anguillicaudatus ). Environmental Toxicology and Pharmacology , 45 : 132-139. (39) Naiel, M.A., Shehata, A.M., Negm, S.S., Abd El‐Hack, M.E., Amer, M.S., Khafaga, A.F., Bin‐Jumah, M. and Allam, A.A. (2020). The new aspects of using some safe feed additives on alleviated imidacloprid toxicity in farmed fish: A review. Reviews in aquaculture , 12 (4): 2250-2267. (40) Qadir, S., Latif, A., Ali, M. and Iqbal, F. (2014). Effects of Imidacloprid on the hematological and serum biochemical profile of Labeo rohita. Pakistan Journal of Zoology, 46 (4): 1085-1090. (41) Topal, A., Alak, G., Ozkaraca, M., Yeltekin, A.C., Comaklı, S., Acıl, G., Kokturk, M. and Atamanalp, M., 2017. Neurotoxic responses in brain tissues of rainbow trout exposed to imidacloprid pesticide: Assessment of 8-hydroxy-2-deoxyguanosine activity, oxidative stress and acetylcholinesterase activity. Chemosphere , 175 : 186-191. (42) Rahman, A.N.A., Mansour, D.A., Abd El-Rahman, G.I., Elseddawy, N.M., Zaglool, A.W., Khamis, T., Mahmoud, S.F. and Mahboub, H.H. (2022). Imidacloprid toxicity in Clarias gariepinus : Protective role of dietary Hyphaene thebaica against biochemical and histopathological disruption, oxidative stress, immune genes expressions, and Aeromonas sobria infection. Aquaculture , 555 : 738170. (43) Farah, A. (1991). Effect of Heavy Metal Toxicants on Liver Function and Hepatic Drug Metabolizing Enzyme System in Rabbit (Doctoral dissertation, University of the Punjab, Lahore). (44) Ali, S.S., Qureshi, M.A., Iqbal, M.J. And Shakoori, A.R. (1992). Zinc-induced biochemical alterations in the liver of common carp, Cyprinus carpio. Punjab University Journal of Zoology, 7 :1-7. (45) Yousafzai, A. M. and Shakoori, A. R. (2011). Hepatic responses of a freshwater fish against aquatic pollution. Pakistan Journal of Zoology, 43 (2): 209-221. (46) Desai, B. and Parikh, P. (2013). Biochemical alterations on exposure of imidacloprid and curzate on fresh water fish Oreochromis mossambicus and Labeo rohita . Indian Journal of Forensic Medicine & Toxicology , 7 (2): 87-91. (47) El Euony, O.I., Elblehi, S.S., Abdel-Latif, H.M., Abdel-Daim, M.M. and El-Sayed, Y.S., 2020. Modulatory role of dietary Thymus vulgaris essential oil and Bacillus subtilis against thiamethoxam-induced hepatorenal damage, oxidative stress, and immunotoxicity in African catfish ( Clarias garipenus ). Environmental Science and Pollution Research , 27 (18): 23108-23128. (48) Rao, V. J. (2006). Sublethal effects of an organophosphorus insecticide (RPR-II) on biochemical parameters of Tilapia, Oreochromis mossambicus . Comparative Biochemistry and Physiology Part C: Toxicology and Pharmacology, 143 (4): 492-498. (49) Sahu, S., Debasmita, S.and Mohapatra, A. K. (2016). Impact of cadmium on some biochemical profiles of the fresh water cat fish Clarias gariepinus (burchell, 1822). The Ecoscan, 9 : 257-265. (50) Sanna, S.G., Tabassum, S., Noreen, S., Mahmood, S., Rehman, M.U., Ahmad, B., Kabir, M., Sajid, M. and Khan, M.F. (2021). Biochemical, Endocrine and Genetic Impairments in Response to Agrochemicals Intoxication in Common Carp (Cyprinus carpio). International Journal of Agriculture & Biology. doi: 10.17957/IJAB/15.1787. (51) Samanta, P., Pal, S., Mukherjee, A.K., Senapati, T. and Ghosh, A.R. (2019). March. Effects of Glyphosate-Based Herbicide (Excel Mera 71) on Digestive Enzymes and Recovery Pattern in Teleostean Fishes. Proceedings of the Zoological Society, 72 (1): 54-60. (52) Samanta, P., Pal, S., Mukherjee, A.K., Senapati, T., Kole, D. and Ghosh, A.R. (2014). Effects of almix herbicide on profile of digestive enzymes of three freshwater teleostean fishes in rice field condition. Toxicology Reports , 1 : 379-384. (53) Samanta, P., Pal, S., Mukherjee, A. K., Senapati, T. and Ghosh, A. R. (2016). Alterations in digestive enzymes of three freshwater teleostean fishes by almix herbicide: A comparative study. Proceedings of the Zoological Society, 69 (1): 61-66. (54) Cornelius, C. E. (1991). Bile Pigments in Fishes: A Review . Veterinary Clinical Pathology, 20 (4): 106–116 . (55) Ujowundu, C. O., Nnanna, C. G., Ndubuisi, E. U., Ngwu, P. C., Uzoma, C. W. and Ezeji, E. U. (2017). Glyphosate-based pesticide-induced biochemical changes in hepatic and renal tissues of Clarias gariepinus . Futo Journal Series, 3 (1): 225-235. (56) Inyang, I. R., Akparanta, O. J. and Izah, S. C. (2019). Evaluation of some blood cells and metabolites of Heterobranchus bidorsalis (hybrid) exposed to sublethal concentrations of sarosate. International Journal of Research in Environmental Science, 5 (1): 17-23. (57) Vitek, L. and Ostrou, J. D. (2009). Bilirubin chemistry and metabolism, harmful and protective aspects. Current Pharmaceutical Design, 15 : 2869-2883. (58) Kumar, V. A., Janaiah, C. and Venkateshwarlu, P. (2010). Effect of thiamethoxam alters serum biochemical parameters in Channa punctatus (Bloch). Asian Journal of Bio Science, 5 (1): 106-110. (59) Nagahama, Y. and Yamashita, M. (2008). Regulation of oocyte maturation in fish. Development, Growth & Differentiation , 50 : S195–S219. (60) Azcoitia, I., Barreto, G.E. and Garcia-Segura, L.M. (2019). Molecular mechanisms and cellular events involved in the neuroprotective actions of estradiol. Analysis of sex differences. Frontiers in Neuroendocrinology , 55 : 100787. (61) Manolagas, S. C. and Kousteni, S. (2001). Perspective: Nonreproductive sites of action of reproductive hormones. Endocrinology , 142 (6): 2200–2204. (62) Bouman, A., Heineman, M. J. and Faas, M. M. (2005). Sex hormones and the immune response in humans. Human Reproduction Update , 111 (4): 411–423. (63) Jin, J., Kurobe, T., Ramírez-Duarte, W. F., Bolotaolo, M. B., Lam, C. H., Pandey, P. K., … Teh, S. J. (2018). Sub-lethal effects of herbicides penoxsulam, imazamox, fluridone and glyphosate on Delta Smelt ( Hypomesus transpacificus ). Aquatic Toxicology, 197 : 79–88. (64) Tripathy, P.S., Parhi, J. and Mandal, S.C., 2021. Steroids and Its Receptors in Fish Reproduction. In Recent updates in molecular Endocrinology and Reproductive Physiology of Fish, (pp. 53-61). Springer, Singapore. (65) Sanderson, C. J. (1992). Interleukin-5, Eosinophils, and Disease. Blood , 79 (12): 3101–3109. (66) Chaves-Pozo, E., García-Ayala, A. and Cabas, I. (2018). Effects of sex steroids on fish leukocytes. Biology , 7 : 9. (67) Chakrabarty, S., Rajakumar, A., Raqhuveer, K., Sridevi, P., Mohanachary, A. and Prathibha Y. (2012). Endosulfan and flutamide, alone and in combination, target ovarian growth in juvenile catfish, Clarias batrachus . Comparative Biochemistry and Physiology, 155 : 491–497. (68) Agbohessi, T.P., Toko, I.I., Atchou, V., Tonato, R., Mandiki, S.N.M. and Kestemont, P. (2015). Pesticides used in cotton production affect reproductive development, endocrine regulation, liver status and offspring fitness in African catfish Clarias gariepinus (Burchell, 1822). Comparative Biochemistry and Physiology, 167 : 157-172 (69) Anjali, V.R., Remya, V.S., Reshmi, S., Mahim, S.S. and Devi, C.A. (2019). Impact of Bisphenol S as an Endocrine Disruptor in a Freshwater Fish, Oreochromis mossambicus . Journal of Endocrinology and Reproduction , 23 (2): 49-63. (70) Marlatt, V.L., Leung, T.Y.G., Calbick, S., Metcalfe, C. and Kennedy, C. (2019). Sub-lethal effects of a neonicotinoid, clothianidin, on wild early life stage sockeye salmon (O ncorhynchus nerka ). Aquatic Toxicology , 217 : 105335. (71) Blanton, M.L. and Specker, J.L. (2007). The hypothalamic-pituitary-thyroid (HPT) axis in fish and its role in fish development and reproduction. Critical Reviews in Toxicology , 37 (1-2): 97-115. (72) Lacasaña, M., López-Flores, I., Rodríguez-Barranco, M., Aguilar-Garduño, C., Blanco-Muñoz, J., Pérez-Méndez, O., Gamboa, R., Bassol, S. and Cebrian, M.E. (2010). Association between organophosphate pesticides exposure and thyroid hormones in floriculture workers. Toxicology and Applied Pharmacology , 243 (1): 19-26. (73) Yu, L., Han, Z. and Liu, C. (2015). A review on the effects of PBDEs on thyroid and reproduction systems in fish. General and Comparative Endocrinology , 219 : 64-73. (74) Campinho, M.A. (2019). Teleost metamorphosis: the role of thyroid hormone. Frontiers in Endocrinology , 10 : 383. (75) Van der Geyten, S., Mol, K.A., Pluymers, W., Kühn, E.R. and Darras, V.M. (1998). Changes in plasma T3 during fasting/refeeding in tilapia ( Oreochromis niloticus ) are mainly regulated through changes in hepatic type II iodothyronine deiodinase. Fish Physiology and Biochemistry , 19 (2): 135-143. (76) Power, D.M., Llewellyn, L., Faustino, M., Nowell, M.A., Björnsson, B.T., Einarsdóttir, I.E., Canario, A.V. and Sweeney, G.E. (2001). Thyroid hormones in growth and development of fish. Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology , 130 (4): 447-459. (77) Ghayyur, S., Khan, M.F., Tabassum, S., Ahmad, M.S., Sajid, M., Badshah, K., Khan, M.A., Ghayyur, S., Khan, N.A., Ahmad, B. and Qamer, S. (2021). A comparative study on the effects of selected pesticides on hemato-biochemistry and tissue histology of freshwater fish Cirrhinus mrigala (Hamilton, 1822). Saudi Journal of Biological Sciences , 28 (1): 603-611. (78) Saha, S., Chukwuka, A.V., Mukherjee, D., Patnaik, L., Nayak, S., Dhara, K., Saha, N.C. and Faggio, C., 2021. Chronic effects of Diazinon® exposures using integrated biomarker responses in freshwater walking catfish, Clarias batrachus. Applied Sciences , 11 (22): 10902. (79) Yu, L.Q., Zhao, G.F., Feng, M., Wen, W., Li, K., Zhang, P.W., Peng, X., Huo, W.J. and Zhou, H.D. (2014). Chronic exposure to pentachlorophenol alters thyroid hormones and thyroid hormone pathway mRNAs in zebrafish. Environmental Toxicology and Chemistry , 33 (1): 170-176. (80) Ndonwi, E.N., Atogho-Tiedeu, B., Lontchi-Yimagou, E., Shinkafi, T.S., Nanfa, D., Balti, E.V., Indusmita, R., Mahmood, A., Katte, J.C., Mbanya, A. and Matsha, T. (2019). Gestational exposure to pesticides induces oxidative stress and lipid peroxidation in offspring that persist at adult age in an animal model. Toxicological Research , 35 (3): 241-248. (81) Abdel-Rahman Mohamed, A., Abdel Rahman, A.N., Salem, G.A., Deib, M.M.E., Nassan, M.A., Rhouma, N.R. and Khater, S.I. (2021). The antioxidant role of a taurine-enriched diet in combating the immunotoxic and inflammatory effects of pyrethroids and/or carbamates in Oreochromis niloticus . Animals , 11 (5): 1318. (82) Borgesa, A.C.P., Piassao, J.F.G., Albani, S.M., Albertoni, E.F., Martins, M.C., Cansian, R.L., Valduga, A.T., Hepp, L.U. and Mielniczki-Pereira, A.A. (2022). Multiple metals and agricultural use affects oxidative stress biomarkers in freshwater Aegla crabs/Presenca de multiplos metais e agricultura afetam biomarcadores de estresse oxidativo em caranguejos de agua doce (Aegla). Brazilian Journal of Biology , 82: e230147. (83) Hellou, J., Ross, N.W. and Moon, T.W. (2012). Glutathione, glutathione S-transferase, and glutathione conjugates, complementary markers of oxidative stress in aquatic biota. Environmental Science and Pollution Research , 19 (6): 2007-2023. (84) Ge, W., Yan, S., Wang, J., Zhu, L., Chen, A. and Wang, J. (2015). Oxidative stress and DNA damage induced by imidacloprid in zebrafish ( Danio rerio ). Journal of agricultural and food chemistry , 63 (6): 1856-1862. (85) John, S., Kale, M., Rathore, N. and Bhatnagar, D. (2001). Protective effect of vitamin E in dimethoate and malathion induced oxidative stress in rat erythrocytes. The Journal of Nutritional Biochemistry , 12 (9): 500-504. (86) Svensson, R., Rinaldi, R., Swedmark, S. and Morgenstern, R. (2000). Reactivity of cysteine-49 and its influence on the activation of microsomal glutathione transferase 1: evidence for subunit interaction. Biochemistry , 39 (49): 15144-15149. (87) Chang, Y., Mao, L., Zhang, L., Zhang, Y. and Jiang, H. (2020). Combined toxicity of imidacloprid, acetochlor, and tebuconazole to zebrafish ( Danio rerio ): acute toxicity and hepatotoxicity assessment. Environmental Science and Pollution Research , 27 (10): 10286-10295. (88) Günal, A.Ç., Erkmen, B., Paçal, E., Arslan, P., Yildirim, Z. and Erkoç, F. (2020). Sub-lethal Effects of Imidacloprid on Nile tilapia ( Oreochromis niloticus ). Water, Air, & Soil Pollution , 231 (1): 1-10. (89) Safi, F., Beger, H.G., Bittner, R., Büchler, M. and Krautzberger, W. (1986). CA 19‐9 and pancreatic adenocarcinoma. Cancer , 57 (4): 779-783. (90) Wahab, M.A., El-Bendary, M., Ali, M.A., Shehta, A., Abed, S., Shehatta, A.S., Salah, M. and El-Gilany, A.H. (2018). Serum Levels of Heavy Metals in Cholangiocarcinoma Patients from the Nile Delta Region of Egypt: A Single Centre Study. Surg. Gastroenterol , 23 (3): 186-190. (91) Abdel-Hamid, O.M., Nafee, A.A., MA, E. and MA, E. (2018). The ameliorative effect of Vitamin C in experimentally induced colon cancer in rats. Benha Veterinary Medical Journal , 34 (1): 329-343. (92) Kim, S., Park, B.K., Seo, J.H., Choi, J., Choi, J.W., Lee, C.K., Chung, J.B., Park, Y. and Kim, D.W. (2020). Carbohydrate antigen 19-9 elevation without evidence of malignant or pancreatobiliary diseases. Scientific reports , 10 : 8820. (93) Sepici-Dinçel, A., Benli, A.Ç.K., Selvi, M., Sarıkaya, R., Şahin, D., Özkul, I.A. and Erkoç, F. (2009). Sublethal cyfluthrin toxicity to carp ( Cyprinus carpio L.) fingerlings: biochemical, hematological, histopathological alterations. Ecotoxicology and Environmental Safety , 72 (5): 1433-1439. (94) Nemeth, E., Baird, A.W. and O’Farrelly, C., 2009, September. Microanatomy of the liver immune system. Seminars in immunopathology, 31 (3): 333-343. (95) Rout, P.C. and Naik, B.N., 1998. Elevation of blood histamine and C-RP durings short-term lead exposure in Clarias batrachus (Linn.). Journal of Nature Conservators , 10 (2): 169-173. (96) Chatterjee, R. (2016). Estimation of endosulfan toxicity on blood biochemistry of channel catfish Clarias batrachus Linn. South Indian Journal of Biological Sciences , 2 (4): 445-450. (97) Paul, I., Mandal, C. and Mandal, C. (1998). Effect of environmental pollutants on the C-reactive protein of a freshwater major carp, Catla catla . Developmental & Comparative Immunology , 22 (5-6): 519-532. (98) Kodama, H., Matsuoka, Y., Tanaka, Y., Liu, Y., Iwasaki, T. and Watarai, S. (2004). Changes of C-reactive protein levels in rainbow trout ( Oncorhynchus mykiss ) sera after exposure to anti-ectoparasitic chemicals used in aquaculture. Fish & shellfish immunology , 16 (5): 589-597. (99) Tarnawska, M., Augustyniak, M., Łaszczyca, P., Migula, P., Irnazarow, I., Krzyżowski, M. and Babczyńska, A. (2019). Immune response of juvenile common carp ( Cyprinus carpio L.) exposed to a mixture of sewage chemicals. Fish and Shellfish Immunology , 88 : 17-27. Supplementary Files Onlinefloatimage1.png Graphical Abstract Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-2193834","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":146285217,"identity":"b7f60a35-4901-4cc9-8c41-438421c39e60","order_by":0,"name":"Nosakhare O. Erhunmwunse","email":"data:image/png;base64,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","orcid":"https://orcid.org/0000-0003-2725-0571","institution":"University of Benin","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Nosakhare","middleName":"O.","lastName":"Erhunmwunse","suffix":""},{"id":146304611,"identity":"cfb42e1d-8440-4e06-9f34-2a0226d93590","order_by":1,"name":"Timeyin J. Pajiah","email":"","orcid":"","institution":"University of Benin","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Timeyin","middleName":"J.","lastName":"Pajiah","suffix":""},{"id":146445260,"identity":"28538146-6bc8-4e87-86fc-dd6bbfb667c9","order_by":2,"name":"Ogbodo Victory","email":"","orcid":"","institution":"University of Benin","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Ogbodo","middleName":"","lastName":"Victory","suffix":""},{"id":146445866,"identity":"4bcbaf78-70e3-4457-ae7d-2a81cfa73fbb","order_by":3,"name":"Endurance Ewere","email":"","orcid":"","institution":"Southern Cross University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Endurance","middleName":"","lastName":"Ewere","suffix":""}],"badges":[],"createdAt":"2022-10-22 15:26:48","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2193834/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2193834/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":28193850,"identity":"b985c71e-3f07-4e8e-abc6-916b3e091a55","added_by":"auto","created_at":"2022-10-24 18:08:56","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":74421,"visible":true,"origin":"","legend":"\u003cp\u003eEffects of imidacloprid on (A) carbohydrate antigen (CA 19-9) level and (B) c-reactive protein (CRP) level in \u003cem\u003eClarias gariepinus\u003c/em\u003e after 96 hours exposure. The results are shown as mean ± standard error of two fish replicates. Asterisk indicates significant difference from control (p \u0026lt; 0.05).\u003c/p\u003e","description":"","filename":"Onlinefloatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-2193834/v1/f601843420ae85539b4a0dd2.png"},{"id":28194213,"identity":"e38abdc0-1995-4435-a719-6c35efc72d09","added_by":"auto","created_at":"2022-10-24 18:14:01","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":643129,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2193834/v1/a1e1beaf-95e8-45f7-9fed-e6c8d3e19def.pdf"},{"id":28194207,"identity":"dfa45c1e-74c9-48b8-a032-e7b667fc64bb","added_by":"auto","created_at":"2022-10-24 18:13:55","extension":"png","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":15943,"visible":true,"origin":"","legend":"\u003cp\u003eGraphical Abstract\u003c/p\u003e","description":"","filename":"Onlinefloatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-2193834/v1/6d459766bc225c4f4701efb9.png"}],"financialInterests":"","formattedTitle":"\u003cp\u003e\u003cstrong\u003eDoes short-term exposure to a Neonicotinoid insecticide trigger Biochemical and Physiological responses in Juvenile catfish?\u003c/strong\u003e\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003ePollution has taken the headlines in recent stories across the globe as more contaminants with varying magnitudes of toxicity are being introduced into aquatic and terrestrial environments. Man's attempt to improve the quality of life subjects the ecological system to enormous stress and deteriorating modifications.\u003csup\u003e1\u003c/sup\u003e These activities affect the health and survival of individual organisms, as well as their performance, and alter the function and structure of natural ecosystems, especially the aquatic ecosystem.\u003csup\u003e2\u003c/sup\u003e Commercial, industrial, and agricultural chemicals can cause several harmful effects on aquatic organisms when they enter the aquatic environment. Fish accumulate these chemicals, including pesticides, directly from the polluted water and indirectly from the food chain.\u003csup\u003e3,4\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eOver a thousand varieties of pesticides are used in the world today, with different properties and exploring diverse modes of action, with more than 80% of farmers relying on pesticides to secure their produce.\u003csup\u003e5\u003c/sup\u003e Some pesticides like DDT and lindane have been proven to be very persistent in soil and water for many years after application.\u003csup\u003e6\u003c/sup\u003e Most pesticides deviate from their main objective of the action, causing a variety of toxic effects in non-target species.\u003csup\u003e7\u003c/sup\u003e As pesticides ultimately flow into the surface water, it is expected that aquatic environments are more affected than terrestrial environments.\u003csup\u003e8,9\u003c/sup\u003e Aquatic ecosystems are threatened by the pollution of these pesticides, thus the development and usage of pesticides with low ecological impact and toxicity to non-target organisms are pertinent for the preservation of these ecosystems and biodiversity.\u003csup\u003e10\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eGreat efforts have been made over the last three decades to produce pesticides that are specific in action, highly degradable, and have selective qualities that work on certain chemical and physiological aspects of insects, reducing the risk of non-target species. This tremendous advance in knowledge paved the way for a more effective group of insecticides, the neonicotinoids, of which imidacloprid is a widely used example. Neonicotinoids are a class of nitroguanidine systemic insecticides often used to treat soil and seeds before planting crops to prevent pest infestations.\u003csup\u003e11\u003c/sup\u003e Owing to their effectiveness against a wide range of insects and the breadth of their uses, as well as their functioning at low dosages with relatively low volatility and high solubility in water.\u003csup\u003e12,13,14\u003c/sup\u003e The idea that neonicotinoids function as nicotinic acetylcholine receptor (nAChR) agonists, which are thought to have little impact on vertebrate species and limited cross-resistance with other pesticides, is the basis for their appeal. Also, neonicotinoids have systemic activity, which allows them to spread more widely throughout plants than conventional pesticides to control the targeted pest species.\u003csup\u003e10\u003c/sup\u003e However, neonicotinoids' high water solubility also makes them very mobile and, as a result, easily transferred into aquatic habitats, potentially causing harm to aquatic organisms that are not intended targets. Seven neonicotinoid pesticides are currently in use, including acetamiprid, dinotefuran, clothianidin, nitenpyram, imidacloprid, thiamethoxam, and thiacloprid.\u003c/p\u003e \u003cp\u003eImidacloprid [1-(6-chloro-3-pyridylmethyl)-N-nitro-imidazolidin-2-ylideneamine] is a neonicotinoid synthetic insecticide that forms an agonistic relationship with the postsynaptic nicotinic acetylcholine receptors\u003csup\u003e15,16\u003c/sup\u003e causing impairment of normal nerve function. It was the first neonicotinoid to be introduced in 1991, and it has since become a key product in several pest control operations.\u003csup\u003e17,18\u003c/sup\u003e Products containing imidacloprid dominate the insecticide industry and are authorized for use in more than 120 countries on more than 140 different types of crops.\u003csup\u003e19\u003c/sup\u003e Its continuously increasing popularity not just in the control of household pets\u0026rsquo; pests but especially in agricultural use can be attributed to its excellent effectiveness in the control of sucking, soil, and chewing insects.\u003csup\u003e20\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eRecently, numerous studies have focused on the toxicity of imidacloprid in the aquatic environment due to the negative effects of neonicotinoids on non-target beneficial invertebrates. Although there is little information available about its toxicity on aquatic vertebrates including fish, several studies have assessed the toxic effects of imidacloprid on several endpoints of different species of fish in various parts of the world.\u003csup\u003e14,21,22\u003c/sup\u003e Imidacloprid has been reported to cause DNA damage and oxidative stress\u003csup\u003e14\u003c/sup\u003e decreased hatching rate of fertilized eggs\u003csup\u003e22\u003c/sup\u003e and histopathological alterations in tissues\u003csup\u003e23\u003c/sup\u003e of different fish species. Nonetheless, no study has conducted a broad range of assessments of behavioral and physiological responses and biomarkers for a fish species. This is necessary to understand the underlying mechanism of action for imidacloprid toxicity in fish and other vertebrates.\u003c/p\u003e \u003cp\u003eConsidering these knowledge gaps, this study aimed to assess the impacts of imidacloprid on \u003cem\u003eClarias gariepinus\u003c/em\u003e behavior, hepatic and endocrine functions, oxidative stress, and tumor and inflammatory responses. To achieve this objective, catfish were exposed to acute concentrations of imidacloprid for 96 hours. \u003cem\u003eC. gariepinus\u003c/em\u003e was chosen for this study due to its wide distribution in tropical freshwater systems, adaptation to laboratory conditions, and stress resistance.\u003csup\u003e24\u003c/sup\u003e\u003c/p\u003e"},{"header":"Materials And Methods","content":"\u003cp\u003e \u003cb\u003eFish Maintenance.\u003c/b\u003e Juvenile \u003cem\u003eClarias gariepinus\u003c/em\u003e (mean length\u0026thinsp;=\u0026thinsp;10.73\u0026thinsp;\u0026plusmn;\u0026thinsp;1.34 cm; mean weight\u0026thinsp;=\u0026thinsp;12.34\u0026thinsp;\u0026plusmn;\u0026thinsp;3.44 g) were purchased from Farm Project, Faculty of Agriculture, University of Benin, Nigeria and maintained in a 160 L plastic tank for two weeks according to OECD\u003csup\u003e25\u003c/sup\u003e at the Laboratory for Ecotoxicology and Environmental Forensics at the University of Benin, Nigeria.\u003c/p\u003e \u003cp\u003e \u003cb\u003eChemical Exposures.\u003c/b\u003e Chemical exposure was conducted through a static renewal system for 96 hours, following OECD guidelines for fish acute toxicity testing\u003csup\u003e25\u003c/sup\u003e. The treatment groups consisted of four concentrations of imidacloprid: 100, 130, 160, and 190 mg/L using a commercially available insecticide, Imi Force\u0026reg; (Jubaili\u0026trade;), containing an imidacloprid concentration of 200 g/L, and a control group. The treatment concentrations were achieved by mixing 10, 13, 16, and 19 mL of the Imi force\u0026reg; insecticide in 20 L water to get the required imidacloprid concentrations of 100, 130, 160, and 190 mg/L respectively. No insecticide or chemical was added to the control group. The concentrations and duration were selected based on previous studies and a range-finding test. Each group was comprised of two replicates (n\u0026thinsp;=\u0026thinsp;6). Mortalities were counted daily for LC\u003csub\u003e50\u003c/sub\u003e determination.\u003c/p\u003e \u003cp\u003e \u003cb\u003eBehavioral Alterations and Mortality.\u003c/b\u003e The fish were inspected multiple times daily during the exposure period to record behavioral changes and clinical diagnostic signs through direct observation, according to OECD\u003csup\u003e25\u003c/sup\u003e. The toxicological endpoint of this study was mortality. Mortality was recorded daily when the fish failed to respond to gentle prodding. The dead fish were removed immediately.\u003c/p\u003e \u003cp\u003e \u003cb\u003eSampling.\u003c/b\u003e All fish were euthanized after 96 hours of exposure to a lethal dose of benzocaine (0.5 g L\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e; Sigma-Aldrich). Samples were selected randomly from each aquarium (n\u0026thinsp;=\u0026thinsp;2), homogenized, and used for the assays.\u003c/p\u003e \u003cp\u003e \u003cb\u003eBiomarker Analysis.\u003c/b\u003e The samples were used in the assay of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities according to Reitman and Frankel\u003csup\u003e26\u003c/sup\u003e; total and conjugate bilirubin levels according to Jendrassik and Grof\u003csup\u003e27\u003c/sup\u003e; amylase activity according to Bern\u003csup\u003e28\u003c/sup\u003e; lipase activity according to Bier\u003csup\u003e29\u003c/sup\u003e; 17β-estradiol (E2) and testosterone levels using an Estradiol EIA Kit (Cayman Chemical, USA) according to the manufacturer\u0026rsquo;s instructions; glutathione S-transferases (GST) activity according to Habig and Jakoby\u003csup\u003e30\u003c/sup\u003e; carbohydrate antigen (CA-19-9) level according to Minamide \u003cem\u003eet al\u003c/em\u003e.\u003csup\u003e31\u003c/sup\u003e; C-reactive protein (CRP) level according to Lowry \u003cem\u003eet al.\u003c/em\u003e\u003csup\u003e32\u003c/sup\u003e; total antioxidant capacity (TAC) activity according to Teimouri \u003cem\u003eet al\u003c/em\u003e.\u003csup\u003e33\u003c/sup\u003e; catalase (CAT) activity according to Beer and Sizer\u003csup\u003e34\u003c/sup\u003e; glutathione peroxidase-1 (GPx) activity according to Hafemann \u003cem\u003eet al.\u003c/em\u003e\u003csup\u003e35\u003c/sup\u003e; superoxide dismutase (SOD) according to McCord and Fridovich\u003csup\u003e36\u003c/sup\u003e; triiodothyronine (T3), thyroxine (T4) and thyroid stimulating hormone (TSH) using ELISA kit (Millipore-Sigma, USA) according to the manufacturer\u0026rsquo;s instructions.\u003c/p\u003e \u003cp\u003e \u003cb\u003eStatistical Analysis.\u003c/b\u003e LC\u003csub\u003e50\u003c/sub\u003e was calculated with probit analysis using Microsoft Excel 2016. Statistical analyses were conducted with GraphPad Prism (version 5.01). All data were analyzed using a single factor one-way analysis of variance (ANOVA), followed by Dunnett\u0026rsquo;s posthoc test. Statistical significance was determined if \u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/p\u003e"},{"header":"Results And Discussion","content":"\u003cp\u003eFish are ideal sentinels for behavioral and health assays of various stressors and toxic chemical exposure due to their constant direct contact with the aquatic environment, where chemical exposure occurs over the entire body surface. Imidacloprid is a promising and rapidly expanding pesticide, known for its selective toxicity to insects. Its detection in food and water prompts the verification of its possible negative effects on non-target organisms. To identify its effect on \u003cem\u003eClarias gariepinus\u003c/em\u003e, various biomarkers were evaluated along with mortality and behavioral changes. This present study contributes to the ongoing efforts to unravel the adverse effects and mode of action of imidacloprid and other neonicotinoid pesticides on non-target species. This study is the first to report behavioral alterations and endocrine disruption in \u003cem\u003eClarias gariepinus\u003c/em\u003e after imidacloprid exposure, as well as the effects of imidacloprid on Ca 19-9 in fish. The results of this study show that exposure to different imidacloprid concentrations adversely impacted\u003cem\u003e\u0026nbsp;C. gariepinus\u003c/em\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eImidacloprid LC\u003csub\u003e50\u003c/sub\u003e, Mortalities, Fish Behavior, and Clinical Symptoms.\u0026nbsp;\u003c/strong\u003eThe water temperature and pH in the aquariums throughout the toxicity testing were 26.64 \u0026plusmn; 0.09 ℃ and 6.74 \u0026plusmn; 0.05 respectively. The survival rate of fish in the control was 100%, indicating that the toxicity data were acceptable.\u003c/p\u003e\n\u003cp\u003eThe mortality rate was time- and concentration-dependent, with % mortality peaking at 66.67% after 96 hours of exposure. Mortality was observed at concentrations \u0026ge;130 mg/L. (Table 1). The 96-h LC\u003csub\u003e50\u003c/sub\u003e of imidacloprid was 166.60 mg/L (95% confidence intervals: 143.38-193.59 mg/L). This quantity is substantially higher than the environmental values already measured and is unlikely to occur in aquatic systems. The value obtained in this study is considerably higher than the previously reported 96-h LC\u003csub\u003e50\u003c/sub\u003e (10.16 \u0026micro;g/L) for \u003cem\u003eC. gariepinus\u003c/em\u003e.\u003csup\u003e21\u003c/sup\u003e However, similar values have been reported for imidacloprid in other fish species, including \u003cem\u003eDanio rerio\u0026nbsp;\u003c/em\u003e(143.7 mg/L)\u003csup\u003e37\u003c/sup\u003e and \u003cem\u003eMisgurnus anguillicaudatus\u003c/em\u003e (145.8 mg/L)\u003csup\u003e38\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:8.0pt;margin-left:0in;line-height:107%;font-size:16px;font-family:\"Times New Roman\",serif;text-align:justify;'\u003e\u003cstrong\u003eTable 1. Cumulative mortality of \u003cem\u003eClarias gariepinus\u0026nbsp;\u003c/em\u003eexposed to various concentrations of imidacloprid\u003c/strong\u003e\u003c/p\u003e\n\u003ctable style=\"border-collapse:collapse;border:none;\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" style=\"width: 85.8pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 26.75pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eImidacloprid concentration (mg/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\" style=\"width: 66.9pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 26.75pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eNumber of exposed fish\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" style=\"width: 257.7pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 26.75pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eCumulative mortality of fish per time interval (h)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd rowspan=\"2\" style=\"width: 64.4pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 26.75pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eMortality\u003c/p\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e%\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 64.4pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 26.75pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 26.75pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 26.75pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 26.75pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e96\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 85.8pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.9pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 85.8pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.9pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 85.8pt;border: medium none;padding: 0in 5.4pt;height: 13.7pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e130\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.9pt;border: medium none;padding: 0in 5.4pt;height: 13.7pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13.7pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13.7pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13.7pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13.7pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13.7pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 85.8pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e160\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.9pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border: medium none;padding: 0in 5.4pt;height: 13pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e42\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 85.8pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e190\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.9pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 64.4pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 12.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e67\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003eFish behavior varied with concentration and time of exposure. All exposed fish showed noticeable distress symptoms, represented by under-reactivity to stimulus, hypoventilation, gulping, abnormal surface distribution, and poor swimming movement (Table 2). Loss of buoyancy was also observed in fish, a few hours before mortality. Fish exposed to \u0026ge; 130 mg/L imidacloprid were severely hypoventilated, exhibiting almost no opercular movement, while fish in \u0026ge; 160 mg/L constantly gasped for air at the water\u0026apos;s surface. This could be a result of imidacloprid affecting fish gills directly, which may affect fish respiration.\u003csup\u003e39\u003c/sup\u003e Although fish activity was largely reduced in all exposed fish compared to the control, fish activity in the 100 mg/L groups showed improvement on the final day of observation. Furthermore, mucus secretion in all exposed fish peaked after 48 hours of exposure but declined after 72 hours. These observations of fish activity and mucus secretion suggest that catfish may recover from acute behavioral effects caused by imidacloprid after a prolonged time of exposure. Literature on the effects of imidacloprid on fish behavior is scarce, however, imidacloprid has been reported to cause similar behavioral alterations by Qadir \u003cem\u003eet al\u003c/em\u003e.\u003csup\u003e40\u003c/sup\u003e in \u003cem\u003eLabeo rohita\u003c/em\u003e exposed to 120 mg/L imidacloprid for 8 days. Qadir \u003cem\u003eet al\u003c/em\u003e.\u003csup\u003e40\u003c/sup\u003e also observed that as exposure time increased, fish tended to recover from the disturbing condition and the occurrence of abnormal behavior declined, similar to our findings. The behavioral changes observed in our study could be attributed to the effect of imidacloprid on the fish nervous system via impairment of brain activity and acetylcholinesterase level alteration, resulting in the impediment of many physiological functions.\u003csup\u003e41,42\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:8.0pt;margin-left:0in;line-height:107%;font-size:16px;font-family:\"Times New Roman\",serif;text-align:justify;'\u003e\u003cstrong\u003eTable 2. Behavioral abnormalities of \u003cem\u003eC. gariepinus\u0026nbsp;\u003c/em\u003edue to the exposure to imidacloprid within the 96-h observation period\u003c/strong\u003e\u003c/p\u003e\n\u003ctable style=\"border-collapse:collapse;border:none;\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eConcentration (mg/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eHypo activity\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eAbnormal surface distribution\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eUnder-reactivity to stimuli\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eHypoventilation\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eGulping\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eMucus secretion\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e24h\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n 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style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e130\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e160\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e190\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n 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top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e130\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e160\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e190\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: 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style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e130\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e160\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e190\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e++\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp 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style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e160\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 78.8pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e190\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.5pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 66.7pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 59.05pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 88.8pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 54.45pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e+++\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 57.5pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e-\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:8.0pt;margin-left:0in;line-height:107%;font-size:16px;font-family:\"Times New Roman\",serif;text-align:justify;'\u003e\u0026apos;-\u0026apos; indicates no behavioral abnormality; \u0026apos;+\u0026apos; indicates the number of individuals showing behavioral abnormalities; \u0026apos;+\u0026apos;= Mild (around 30 to 40% of individuals showing behavioral abnormalities); \u0026apos;++\u0026apos; = Moderate (around 45 to 65% of individuals showing the behavioral abnormalities); \u0026lsquo;+++\u0026rsquo; = Severe (\u0026ge; 70% of individuals showing the behavioral abnormalities).\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eHepatic Biomarkers.\u0026nbsp;\u003c/strong\u003eConcentrations of ALT and AST were increased in fish exposed to imidacloprid, however, these changes were not significantly different from the control. ALT levels increased with increasing concentrations of imidacloprid until 160 mg/L peaking at a 176% difference from the control. AST levels increased considerably in fish exposed to imidacloprid, with a difference of more than 17 folds observed at 130 mg/L (Table 3). The increased enzymatic activities could result from elevated enzyme production to counter the harm caused by the insecticide or a disturbance in the cell membrane integrity, preventing the enzymes from moving freely.\u003csup\u003e43,44,45\u003c/sup\u003e Increases in ALT and AST levels have previously been reported in \u003cem\u003eC. gariepinus\u003c/em\u003e and other fish species exposed to neonicotinoids.\u003csup\u003e40,46,47\u003c/sup\u003e Desai and Parikh\u003csup\u003e46\u003c/sup\u003e studied the biochemical effects of imidacloprid on \u003cem\u003eLabeo rohita\u0026nbsp;\u003c/em\u003eand \u003cem\u003eOreochromis mossambicus\u003c/em\u003e. Fish exposure to imidacloprid for 21 days elevated AST and ALT levels in several tissues of \u003cem\u003eOreochromis mossambicus\u003c/em\u003e and \u003cem\u003eLabeo rohita\u003c/em\u003e.\u003csup\u003e46\u003c/sup\u003e Increased activities of ALT and AST could be due to amplified transamination processes that occur as a result of amino acid input into the tricarboxylic acid cycle to manage energy crises during toxicant-based stress.\u003csup\u003e48,49\u003c/sup\u003e The likelihood of the elevation of the enzymes due to an energy crisis is supported by our observations on amylase and lipase activity in exposed groups.\u003c/p\u003e\n\u003cp\u003eAmylase activity in catfish increased with increasing concentrations of imidacloprid. A significant increase in amylase activity was observed in catfish exposed to 190 mg/L imidacloprid. Similarly, a concentration-dependent increase was observed in lipase activity. A significant difference from the control was observed in fish exposed to \u0026ge; 130 mg/L with the confidence intervals increasing with each observed concentration of imidacloprid (Table 3). Amylase and lipase are important enzymes that are directly related to carbohydrate and lipid metabolism. Amylase is secreted by the exocrine region of the pancreas. The elevated activity could be the result of amylase secretory cell damage or pancreatitis.\u003csup\u003e45\u003c/sup\u003e Alteration in serum amylase indicates an abnormality of the digestive process.\u003csup\u003e50\u003c/sup\u003e In biological systems, lipase is crucial for maintaining the integrity of membranes and cell signaling.\u003csup\u003e51\u003c/sup\u003e Increased lipase activity could be due to tissue damage, the development of a stress situation, or the effective use of lipid molecules for digestion to match the high energy demand during the imidacloprid-induced stress condition.\u003csup\u003e52\u003c/sup\u003e Literature available on the effects of imidacloprid on lipase and amylase activities in fish is limited, however, increases in their activities in several fish species due to other pesticides have been reported.\u003csup\u003e50,51,52,53\u003c/sup\u003e Our study indicates that imidacloprid can impact fish metabolic activities, forcing increased energy production through carbohydrate and lipid metabolism.\u003c/p\u003e\n\u003cp\u003eNo significant difference was observed between total bilirubin levels in control and catfish exposed to imidacloprid. The total bilirubin levels in exposed catfish were generally higher than the control, with a drop below the control level observed only in the 190 mg/L groups. In contrast, a significant difference from the control was observed in conjugate bilirubin levels of the fish exposed to 130 mg/L. However, similar to the patterns observed in total bilirubin levels, conjugate bilirubin levels of catfish exposed to imidacloprid were generally higher than that of the control, with a drop below control levels observed only in the 190 mg/L groups (Table 3). Bilirubin is one of the main bile pigments found in the fish circulatory system and is derived from hemoglobin disruption.\u003csup\u003e54\u003c/sup\u003e The increased bilirubin production could result from imidacloprid inducing increased red blood cell breakdown or reduced hepatic uptake of bilirubin due to the decreased transport protein-albumin level\u003csup\u003e55\u003c/sup\u003e, severe damage to hepatocytes, and hindrance of the bile duct.\u003csup\u003e56\u003c/sup\u003e Increase in total and conjugate bilirubin levels have also been linked to liver and biliary tract disease.\u003csup\u003e57\u003c/sup\u003e Experimental studies have shown that exposure of \u003cem\u003eC. gariepinus\u0026nbsp;\u003c/em\u003eand other fish species to neonicotinoids, including imidacloprid, causes an increase in bilirubin levels\u003csup\u003e40,58\u003c/sup\u003e which among others has been attributed to acute hepatic necrosis.\u003csup\u003e58\u003c/sup\u003e\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:8.0pt;margin-left:0in;line-height:107%;font-size:16px;font-family:\"Times New Roman\",serif;text-align:justify;'\u003e\u003cstrong\u003eTable 3. Effects on hepatic functions of \u003cem\u003eC. gariepinus\u0026nbsp;\u003c/em\u003eafter 96 hours of exposure to imidacloprid\u003c/strong\u003e\u003c/p\u003e\n\u003ctable style=\"width:538.65pt;margin-left:-22.95pt;border-collapse:collapse;border:none;\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 72.8pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eImidacloprid (mg/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 67.9pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eALT (U/l)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 86.15pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eAST (U/l)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.95pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eAmylase (U/l)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 85.05pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eLipase (U/l)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 70.9pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eTotal bilirubin (\u0026micro;mol/l)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.9pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eConjugate bilirubin (\u0026micro;mol/l)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 72.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 67.9pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e3.38 \u0026plusmn; 1.24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 86.15pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e4.45 \u0026plusmn; 0.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.95pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e3.70 \u0026plusmn; 1.60\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 85.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.15 \u0026plusmn; 0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 70.9pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.31 \u0026plusmn; 0.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.9pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.06 \u0026plusmn; 0.02\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 72.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 67.9pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e7.51 \u0026plusmn; 3.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 86.15pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e11.75 \u0026plusmn; 1.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.95pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e7.95 \u0026plusmn; 0.55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 85.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.44 \u0026plusmn; 0.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 70.9pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.37 \u0026plusmn; 0.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.9pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.14 \u0026plusmn; 0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 72.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e130\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 67.9pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e8.58 \u0026plusmn; 2.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 86.15pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e78.22 \u0026plusmn; 18.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.95pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e10.05 \u0026plusmn; 4.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 85.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.55 \u0026plusmn; 0.03*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 70.9pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.58 \u0026plusmn; 0.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.9pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.28 \u0026plusmn; 0.07*\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 72.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e160\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 67.9pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e9.35 \u0026plusmn; 2.92\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 86.15pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e67.50 \u0026plusmn; 20.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.95pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e17.45 \u0026plusmn; 4.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 85.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.70 \u0026plusmn; 0.03**\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 70.9pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.37 \u0026plusmn; 0.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.9pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.18 \u0026plusmn; 0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 72.8pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e190\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 67.9pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e6.43 \u0026plusmn; 0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 86.15pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e66.45 \u0026plusmn; 21.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.95pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e23.80 \u0026plusmn; 1.60*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 85.05pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e1.13 \u0026plusmn; 0.01***\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 70.9pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.12 \u0026plusmn; 0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.9pt;border: medium none;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.06 \u0026plusmn; 0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 72.8pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e\u003cem\u003eP\u0026nbsp;\u003c/em\u003evalue\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 67.9pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.4537\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 86.15pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.0541\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.95pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.0340\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 85.05pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.0013\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 70.9pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.4383\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.9pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.0189\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:8.0pt;margin-left:0in;line-height:107%;font-size:16px;font-family:\"Times New Roman\",serif;text-align:justify;'\u003eThe values are mean \u0026plusmn; standard error of two fish replicates. *, ** and *** indicate significant difference between exposure and control group at p \u0026lt; 0.05, 0.01 and 0.001 respectively.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEndocrine Disrupting Biomarkers.\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eCatfish 17\u0026beta;-estradiol (E2) levels increased with increasing concentrations of imidacloprid, having an 85% increase from control levels in the group exposed to 190 mg/L. However, these increments were not significantly different from the control. Testosterone levels in catfish fluctuated minimally with increasing concentrations of imidacloprid, with a maximum 5% difference from the levels observed in the control fish. These changes were not significantly different from the control as well (Table 4). The primary internal regulators of gonadal differentiation and its activities are sex steroid hormones.\u003csup\u003e59\u003c/sup\u003e They influence how the brain is organized during the formative stages and sex-dependently coordinate a range of behaviors.\u003csup\u003e60\u003c/sup\u003e They also have an impact on the neurological, skeletal, and cardiovascular systems, as well as stress and immunological responses.\u003csup\u003e61,62\u003c/sup\u003e E2 is a major sex hormone that controls female fish maturity, and high E2 levels are a reliable sign of endocrine disturbance.\u003csup\u003e63\u003c/sup\u003e Elevated E2 concentrations are especially worrisome because they can upset the endocrine system\u0026apos;s delicate balance, impairing reproductive function and resulting in lower fecundity and fertility, repressed sexual behavior, and altered sexual differentiation.\u003csup\u003e63\u003c/sup\u003e Fish immune systems and numerous leukocyte functions are regulated by estrogens.\u003csup\u003e64\u003c/sup\u003e Acidophilic granulocytes, which have been linked to tissue damage\u003csup\u003e65\u003c/sup\u003e, migrate from the head kidney as a result of an increase in E2 levels following an inflammatory reaction.\u003csup\u003e66\u003c/sup\u003e This is the first study on the effects of imidacloprid on catfish sex steroid hormones. Experimental studies have shown that \u003cem\u003eClarias\u0026nbsp;\u003c/em\u003eand several fish species\u0026apos; exposure to pesticides and other stressors cause elevations in E2 levels.\u003csup\u003e63,67,68,69,70\u003c/sup\u003e Additional research is necessary to identify whether imidacloprid affects E2 levels directly or indirectly through the conversion of testosterone to E2, slowed sex steroid hormone degradation mechanisms, or at the hypothalamic-pituitary-gonad axis. The findings indicate that additional research is required to fully understand the negative effects of imidacloprid on the reproductive endocrine axis, regardless of the mode of action that led to the elevated E2 reported in this study.\u003c/p\u003e\n\u003cp\u003eSignificant differences were observed between T3 levels in the control and catfish exposed to 190 mg/L. T3 and T4 levels were generally reduced with increasing concentrations of imidacloprid. However, no significant difference was observed between T4 levels in control and fish exposed to imidacloprid. In contrast, TSH levels of fish increased on exposure to imidacloprid, with a significant difference observed between the control and 190 mg/L groups (Table 4). Thyroid hormones in fish regulate growth, development, metabolism, skin pigmentation, behavior, metamorphosis, osmoregulation, and lipid and protein metabolism, often in association with growth hormone and cortisol.\u003csup\u003e71,72,73,74\u003c/sup\u003e Production of T3 is largely by peripheral enzymatic monodeiodination of T4 majorly in the liver and other tissues.\u003csup\u003e75\u003c/sup\u003e Typically, a decrease in T4 synthesis and secretion is what causes T3 levels to decrease.\u003csup\u003e71\u003c/sup\u003e In the present study, a decrease in the level of thyroid hormones might be indicative of metabolic energy redirecting away from the anabolic process, which is essential to carry through life, thereby triggering thyroid endocrine disruption.\u003csup\u003e76\u003c/sup\u003e Thyroid hormone levels are regulated by the negative feedback that thyroid hormones have on their upstream regulators. Reductions in T3 or T4 enhance the production of TSH by the pituitary gland, hence our findings could be a reflection of this mechanism. Ghayyur \u003cem\u003eet al\u003c/em\u003e.\u003csup\u003e77\u003c/sup\u003e reported similar findings in their 30-day exposure of \u003cem\u003eCirrhinus mrigala\u003c/em\u003e to the neonicotinoid acetamiprid. They observed significant increases in thyroid hormones and a significant reduction in TSH levels in their experiment after fish exposure to acetamiprid, chlorfenapyr, and dimethoate. Saha \u003cem\u003eet al\u003c/em\u003e.\u003csup\u003e78\u003c/sup\u003e observed a reduction in thyroid hormone levels in \u003cem\u003eClarias batrachus\u003c/em\u003e after chronic exposure to diazinon and suggested these reductions could be due to pesticide effects on thyroid hormone signaling and metabolism in the hypothalamic-pituitary-thyroid axis.\u003csup\u003e78,79\u003c/sup\u003e Our study highlights the disruptive endocrine properties of imidacloprid, confirming its potential to adversely affect normal ecological functions and the survival of fish.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:8.0pt;margin-left:0in;line-height:107%;font-size:16px;font-family:\"Times New Roman\",serif;text-align:justify;'\u003e\u003cstrong\u003eTable 4. Effects on endocrine functions of \u003cem\u003eC. gariepinus\u0026nbsp;\u003c/em\u003eafter 96 hours of exposure to imidacloprid\u003c/strong\u003e\u003c/p\u003e\n\u003ctable style=\"width:502.75pt;border-collapse:collapse;border:none;\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 79.1pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 34pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eImidacloprid (mg/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 96.4pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 34pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e17\u0026beta;-estradiol (ng/ml)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 79.05pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 34pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eTestosterone (ng/ml)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 92.8pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 34pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eT3 (ng/ml)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.2pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 34pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eT4 (\u0026micro;g/dL)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 78.2pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 34pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eTSH (\u0026micro;lm/mL)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 79.1pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 96.4pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e68.60 \u0026plusmn; 4.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 79.05pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.80 \u0026plusmn; 0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 92.8pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e1.05 \u0026plusmn; 0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.2pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e7.40 \u0026plusmn; 1.30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 78.2pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.70 \u0026plusmn; 0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 79.1pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 96.4pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e106.30 \u0026plusmn; 15.95\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 79.05pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.80 \u0026plusmn; 0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 92.8pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e1.00 \u0026plusmn; 0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.2pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e6.50 \u0026plusmn; 0.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 78.2pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.80 \u0026plusmn; 0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 79.1pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e130\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 96.4pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e108.70 \u0026plusmn; 16.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 79.05pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.84 \u0026plusmn; 0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 92.8pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e1.05 \u0026plusmn; 0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.2pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e5.45 \u0026plusmn; 0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 78.2pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.80 \u0026plusmn; 0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 79.1pt;border: medium none;padding: 0in 5.4pt;height: 12.55pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e160\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 96.4pt;border: medium none;padding: 0in 5.4pt;height: 12.55pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e111.50 \u0026plusmn; 5.65\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 79.05pt;border: medium none;padding: 0in 5.4pt;height: 12.55pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.81 \u0026plusmn; 0.02\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 92.8pt;border: medium none;padding: 0in 5.4pt;height: 12.55pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.90 \u0026plusmn; 0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.2pt;border: medium none;padding: 0in 5.4pt;height: 12.55pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e5.35 \u0026plusmn; 0.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 78.2pt;border: medium none;padding: 0in 5.4pt;height: 12.55pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.80 \u0026plusmn; 0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 79.1pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e190\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 96.4pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e126.60 \u0026plusmn; 17.40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 79.05pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.78 \u0026plusmn; 0.01\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 92.8pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.85 \u0026plusmn; 0.05*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.2pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e4.35 \u0026plusmn; 0.35\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 78.2pt;border: medium none;padding: 0in 5.4pt;height: 11.9pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.95 \u0026plusmn; 0.05**\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 79.1pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 11.25pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e\u003cem\u003eP\u0026nbsp;\u003c/em\u003evalue\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 96.4pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 11.25pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.1561\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 79.05pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 11.25pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.0551\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 92.8pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 11.25pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.0448\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 77.2pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 11.25pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.2019\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 78.2pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 11.25pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.0047\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:8.0pt;margin-left:0in;line-height:107%;font-size:16px;font-family:\"Times New Roman\",serif;text-align:justify;'\u003eThe values are mean \u0026plusmn; standard error of two fish replicates. * and ** indicate a significant difference between the exposure and control group at p \u0026lt; 0.05 and 0.01 respectively.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eOxidative Biomarkers.\u0026nbsp;\u003c/strong\u003eSignificant reductions were observed between SOD activity in control and exposed fish at different concentrations of imidacloprid. GPx activity of fish increased in groups exposed to \u0026ge; 130 mg/L imidacloprid. A significant difference between GPx activity in control and exposed fish was observed in 130 mg/L imidacloprid concentration. A non-significant increase in CAT, TAC, and GST activity of the exposed groups compared to the control group was observed (Table 5). Pesticide exposure has the potential to cause oxidative damage.\u003csup\u003e80\u003c/sup\u003e To deal with oxidative stress and prevent excessive reactive oxygen species (ROS) formation, which can cause oxidative damage to nucleic acids, lipids, and proteins, fish produce a variety of anti-oxidative defenses including GPx, CAT, and SOD.\u003csup\u003e21,41,81,82\u003c/sup\u003e The role of GST is to catalyze the detoxification of harmful compounds via the conjugation of glutathione.\u003csup\u003e83\u003c/sup\u003e The current study demonstrated reduced SOD, and elevated GPx, CAT, GST, and TAC in \u003cem\u003eC. gariepinus\u003c/em\u003e exposed to imidacloprid. An increase in CAT, GPx, and TAC activity could be an indication that imidacloprid caused an overproduction of ROS in exposed fish.\u003csup\u003e14\u003c/sup\u003e The reduced SOD activity may be due to the elevated oxygen free radicals in catfish, rendering SOD inactive through oxidation.\u003csup\u003e84\u003c/sup\u003e Additionally, elevated quantities of H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e and superoxide anions may be the cause of the rise in CAT activity.\u003csup\u003e85\u003c/sup\u003e The increase in H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e levels is supported by Svensson \u003cem\u003eet al\u003c/em\u003e.\u003csup\u003e86\u003c/sup\u003e who reported that the sulfhydryl of microsomal GST\u0026rsquo;s 49th cysteine could specifically combine with H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e, thus enhancing GST activity. Nnadi \u003cem\u003eet al\u003c/em\u003e.\u003csup\u003e21\u003c/sup\u003e reported significant SOD activity reductions and CAT activity elevations in \u003cem\u003eC. gariepinus\u0026nbsp;\u003c/em\u003eexposed to imidacloprid, similar to our findings. However, contrary to our results, they found that imidacloprid induced a significant reduction in GPx activity in exposed catfish. Several studies have reported oxidative stress induced by imidacloprid in various fish species such as \u003cem\u003eDanio rerio\u003c/em\u003e\u003csup\u003e87\u003c/sup\u003e, \u003cem\u003eOncorhynchus mykiss\u003c/em\u003e\u003csup\u003e41\u003c/sup\u003e, \u003cem\u003eOreochromis niloticus\u003c/em\u003e\u003csup\u003e88\u003c/sup\u003e, and \u003cem\u003eProchilodus lineatus\u003c/em\u003e\u003csup\u003e14\u003c/sup\u003e. Our study further affirms the potential of imidacloprid to induce oxidative stress in fish.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:8.0pt;margin-left:0in;line-height:107%;font-size:16px;font-family:\"Times New Roman\",serif;text-align:justify;'\u003e\u003cstrong\u003eTable 5. Effects on oxidative stress of \u003cem\u003eC. gariepinus\u0026nbsp;\u003c/em\u003eafter 96 hours of exposure to imidacloprid\u003c/strong\u003e\u003c/p\u003e\n\u003ctable style=\"width:491.75pt;border-collapse:collapse;border:none;\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 82.5pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 20.65pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eImidacloprid (mg/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.85pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 20.65pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eSOD (U/ml)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 90.05pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 20.65pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eGPx (Unit/ml)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 84.8pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 20.65pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eCAT (mmol/min/ml)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75.75pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 20.65pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eTAC (mmol/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 84.8pt;border-color: windowtext currentcolor;border-style: solid none;border-width: 1pt medium;padding: 0in 5.4pt;height: 20.65pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003eGST (mmol/mi/min)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 82.5pt;border: medium none;padding: 0in 5.4pt;height: 10.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.85pt;border: medium none;padding: 0in 5.4pt;height: 10.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e1.81 \u0026plusmn; 0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 90.05pt;border: medium none;padding: 0in 5.4pt;height: 10.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e11.20 \u0026plusmn; 2.80\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 84.8pt;border: medium none;padding: 0in 5.4pt;height: 10.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e22.30 \u0026plusmn; 3.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75.75pt;border: medium none;padding: 0in 5.4pt;height: 10.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e1.59 \u0026plusmn; 0.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 84.8pt;border: medium none;padding: 0in 5.4pt;height: 10.85pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.55 \u0026plusmn; 0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 82.5pt;border: medium none;padding: 0in 5.4pt;height: 10.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e100\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.85pt;border: medium none;padding: 0in 5.4pt;height: 10.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e1.44 \u0026plusmn; 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0.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 90.05pt;border: medium none;padding: 0in 5.4pt;height: 10.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e33.65 \u0026plusmn; 8.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 84.8pt;border: medium none;padding: 0in 5.4pt;height: 10.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e40.35 \u0026plusmn; 3.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75.75pt;border: medium none;padding: 0in 5.4pt;height: 10.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e1.81 \u0026plusmn; 0.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 84.8pt;border: medium none;padding: 0in 5.4pt;height: 10.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e3.15 \u0026plusmn; 2.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd style=\"width: 82.5pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 10.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e\u003cem\u003eP\u0026nbsp;\u003c/em\u003evalue\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 73.85pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 10.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.0244\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 90.05pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 10.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.0353\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 84.8pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 10.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.3369\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 75.75pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 10.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.3080\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd style=\"width: 84.8pt;border-color: currentcolor currentcolor windowtext;border-style: none none solid;border-width: medium medium 1pt;border-image: none 100% / 1 / 0 stretch;padding: 0in 5.4pt;height: 10.3pt;vertical-align: top;\"\u003e\n \u003cp style='margin-top:0in;margin-right:0in;margin-bottom:0in;margin-left:0in;line-height:normal;font-size:16px;font-family:\"Times New Roman\",serif;text-align:center;'\u003e0.5814\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp style='margin-top:0in;margin-right:0in;margin-bottom:8.0pt;margin-left:0in;line-height:107%;font-size:16px;font-family:\"Times New Roman\",serif;text-align:justify;'\u003eThe values are mean \u0026plusmn; standard error of two fish replicates. * indicates a significant difference between the exposure and control group at p \u0026lt; 0.05.\u003c/p\u003e\n\u003cp\u003e\u003cbr\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eTumor and Inflammatory Biomarkers\u003c/strong\u003e. Significant increases (\u003cem\u003ep\u0026nbsp;\u003c/em\u003e= 0.0270) between CA 19-9 levels in control and fish exposed to imidacloprid were observed at imidacloprid concentrations \u0026ge; 160mg/L (Figure 1A). Increases in serum CA 19-9 are helpful in the diagnosis of upper gastrointestinal tract adenocarcinoma and the monitoring of colonic carcinoma, but their greatest sensitivity has been found in the detection of pancreatic adenocarcinoma, according to various studies.\u003csup\u003e89,90\u003c/sup\u003e This is the first study assessing pesticide effects on fish CA 19-9 levels. Literature on the effects of toxicants on CA 19-9 in animal studies is incredibly scarce, however, Omnia \u003cem\u003eet al\u003c/em\u003e.\u003csup\u003e91\u003c/sup\u003e observed a significant increase in CA 19-9 levels in rats exposed to 1,2-dimethylhydrazine, similar to our results. Our findings indicate that imidacloprid may pose a high risk of malignant and pancreaticobiliary diseases in fish. However, Kim \u003cem\u003eet al\u003c/em\u003e.\u003csup\u003e92\u003c/sup\u003e found CA-19-9 elevation in patients with pulmonary, gynecologic, endocrine, and splenic diseases who did not have a malignant and pancreaticobiliary disease.\u003c/p\u003e\n\u003cp\u003eThere was no significant difference (\u003cem\u003ep\u0026nbsp;\u003c/em\u003e= 0.0807) between CRP levels in the control and imidacloprid-exposed groups. Furthermore, the CRP levels fluctuated, and the comparison of these two groups did not yield a consistent pattern over time, having only minor deviations from the control group levels except in the group exposed to 190 mg/L (Figure 1B). As an initial line of defense against environmental contaminants, toxic chemicals cause fish to produce more non-antibody proteins like CRP.\u003csup\u003e93\u003c/sup\u003e CRP is mostly produced in the liver and is responsible for several immunological responses.\u003csup\u003e94\u003c/sup\u003e Our studies reveal that although imidacloprid has a mild effect on CRP levels at concentrations below the 96-h LC\u003csub\u003e50\u003c/sub\u003e, its effects become more noticeable at higher concentrations, where a 32% increase in CRP levels was observed in catfish exposed to 190 mg/L imidacloprid. Currently, there is no comprehensive data available on CRP levels due to imidacloprid and other neonicotinoid exposure in fish with which to compare this data. However, elevated CRP levels have been observed in \u003cem\u003eClarias\u003c/em\u003e species on exposure to various stressors.\u003csup\u003e95,96\u003c/sup\u003e Chatterjee\u003csup\u003e96\u003c/sup\u003e exposed \u003cem\u003eC. gariepinus\u0026nbsp;\u003c/em\u003eto endosulfan for 96 hours and observed a steady rise in CRP levels. Furthermore, pesticides, as well as other physiological and chemical stressors, have been reported to increase CRP levels in several fish species.\u003csup\u003e93,97,98,99\u003c/sup\u003e This elevated level of CRP in the acute phase response indicates that fish exposed to imidacloprid may develop inflammatory responses.\u003c/p\u003e\n\u003cp\u003eIn conclusion, short-term exposure to imidacloprid, a neonicotinoid insecticide, now largely used worldwide as a replacement for more toxic and persistent pesticides triggered biochemical and physiological responses in juvenile catfish (\u003cem\u003eClarias gariepinus\u003c/em\u003e). The present study clearly showed that imidacloprid has the potential to disrupt various physiological and behavioral functions in fish, which could lower the probability of survival. While extensive research has focused on the effects of imidacloprid on fish behavior, transaminases, and oxidative stress, our work highlights the need for further examination of its effects on hepatic and endocrine functions, as well as tumor and inflammatory responses. Whether or not the environmentally relevant concentrations of imidacloprid will cause these same effects in catfish remains yet to be known. Controlled use of imidacloprid-based pesticides, especially close to aquatic ecosystems, is therefore essential.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eEthical approval for this research was given by the Research Ethics committee of the College of Medicine of the University of Benin. \u003cstrong\u003eCMS/REC/2020/34.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and material\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCRediT authorship contribution statement\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNosakhare O. Erhunmwunse: \u003cstrong\u003eConceived and designed the experiments\u003c/strong\u003e. Timeyin J. Pajiah, Junior V. Ogodo\u003cstrong\u003e\u0026nbsp;performed the experiments\u003c/strong\u003e. Nosakhare O. Erhunmwunse, Timeyin J. Pajiah, Junior V. Ogodo: \u003cstrong\u003eAnalyzed and interpreted the data\u003c/strong\u003e. Nosakhare O. Erhunmwunse, Timeyin J. Pajiah, Junior V. Ogodo, Endurance Ewere: \u003cstrong\u003eMaterials, analysis tools, and ideas\u003c/strong\u003e. All authors have read and approved the manuscripts.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeclaration of Competing Interest\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgment\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe appreciate the diligence, and relentless efforts of Mr. Christopher who helped us all through the work.\u003c/p\u003e"},{"header":"References","content":"\u003cp\u003e(1) Lopez-Barea, J. (1995). Biomarkers in ecotoxicology\u003cem\u003e. Archives of toxicology,\u003c/em\u003e\u003cstrong\u003e\u0026nbsp;17\u003c/strong\u003e: 57-79.\u003c/p\u003e\n\u003cp\u003e(2) Yancheva, V., Velcheva, I., Stoyanova, S and Georgieva, E. (2016). Histological biomarkers in fish as a tool in ecological risk assessment and monitoring programs: A review. \u003cem\u003eApplied Ecology and Environment Research,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e14\u003c/strong\u003e(1): 47-75.\u003c/p\u003e\n\u003cp\u003e(3) Mohamed, A. S. F. (2009). Histopathological studies on \u003cem\u003eTillapia zilli\u003c/em\u003e and \u003cem\u003eSolea vulgaris\u003c/em\u003e from Lake Qarun, Egypt. \u003cem\u003eWorld Journal of Fish and Marine Sciences,\u003c/em\u003e \u003cstrong\u003e1\u003c/strong\u003e: 29-39.\u003c/p\u003e\n\u003cp\u003e(4) Chaudhry, A.S., and Jabeen, F. (2011). Assessing metal, protein and DNA profiles in \u003cem\u003eLabeo rohita\u003c/em\u003e from the Indus River in Mianwali, Pakistan\u003cem\u003e. Environmental Monitoring and Assessment\u003c/em\u003e, \u003cstrong\u003e174\u003c/strong\u003e: 665-679.\u003c/p\u003e\n\u003cp\u003e(5) Shahzad, F., Taj, M. K., Abbas, F., Taj, I., Parveen, S., Achakzai, A. M., Azam, S., Hussain, A., Tareen, A. R., Mohammad, G., Samreen, Z., Sazian, B. and Bibi, L. (2019). Pesticides and our environment. \u003cem\u003eInternational Journal of Biosciences,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e14\u003c/strong\u003e(4): 487-491.\u003c/p\u003e\n\u003cp\u003e(6) Hadian, Z., Eslamizad, S. and Yazdanpanah, H. (2019). Pesticide Residues Analysis in Iranian Fruits and Vegetables by Gas Chromatography-Mass Spectrometry. \u003cem\u003eIranian Journal of Pharmaceutical Research\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e18\u003c/strong\u003e: 275.\u003c/p\u003e\n\u003cp\u003e(7) Maton, S.M., Dodo, J.D., Nesla, R.A. and Ali, A.Y. (2016). Environmental Impact of Pesticides Usage on Farmlands in Nigeria. \u003cem\u003eInternational journal of innovative research and development\u003c/em\u003e \u003cstrong\u003e5\u003c/strong\u003e: 311-317.\u003c/p\u003e\n\u003cp\u003e(8) Pritchard, J. B. (1993). Aquatic toxicology: past, present, and prospects. \u003cem\u003eEnvironmental Health Perspectives,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e100\u003c/strong\u003e: 249\u0026ndash;257.\u003c/p\u003e\n\u003cp\u003e(9) Al-Badran, A. A., Fujiwara, M. and Mora, M. A. (2019). Effects of insecticides, fipronil and imidacloprid, on the growth, survival, and behavior of brown shrimp \u003cem\u003eFarfantepenaeus aztecus\u003c/em\u003e. \u003cem\u003ePLoS ONE,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e14\u003c/strong\u003e(10):e0223641.\u003c/p\u003e\n\u003cp\u003e(10) Hayasaka, D., Suzuki, K., Nomura, T., Nishiyama, M., Nagai, T., S\u0026aacute;nchez-Bayo, F. and Goka, K. (2013). Comparison of acute toxicity of two neonicotinoid insecticides, imidacloprid and clothianidin, to five cladoceran species. \u003cem\u003eJournal of Pesticide Science\u003c/em\u003e, \u003cstrong\u003e38\u003c/strong\u003e(1): 44\u0026ndash;47.\u003c/p\u003e\n\u003cp\u003e(11) Morrissey, C. A., Mineau, P., Devries, J. H., Sanchez-Bayo, F., Liess, M., Cavallaro, M. C. and Liber, K. (2015). Neonicotinoid contamination of global surface waters and associated risk to aquatic invertebrates: A review. \u003cem\u003eEnvironment International,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e74\u003c/strong\u003e: 291-303.\u003c/p\u003e\n\u003cp\u003e(12) EXTOXNET, (1996). Extension Toxicology Network. Pesticide Information profiles: Imidacloprid. Available at http://extoxnet.orst.edu/pips/imidaclo.htm, accessed on 13 April 2022.\u003c/p\u003e\n\u003cp\u003e(13) Tomlin, C. D. S. (2000). The Pesticide Manual, twelfth ed. British Crop Protection Council, Surrey, United Kingdom.\u003c/p\u003e\n\u003cp\u003e(14) Vieira, C. E. D., P\u0026eacute;rez, M. R., Acayaba, R. D., Raimundo, C. C. M. and Martinez, C. B. R. (2018). DNA damage and oxidative stress induced by imidacloprid exposure in different tissues of the Neotropical fish \u003cem\u003eProchilodus lineatus\u003c/em\u003e. \u003cem\u003eChemosphere,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e195\u003c/strong\u003e: 125-134.\u003c/p\u003e\n\u003cp\u003e(15) Radwan, M. A. and Mohamed, M. S. (2013). Imidacloprid induced alterations in enzyme activities and energy reserves of the land snail, \u003cem\u003eHelix aspersa. Ecotoxicology and Environmental Safety,\u003c/em\u003e \u003cstrong\u003e95\u003c/strong\u003e: 91-97.\u003c/p\u003e\n\u003cp\u003e(16) Shan, Y., Yan, S., Hong, X., Zha, J. and Qin, J. (2020). Effect of imidacloprid on the behavior, antioxidant system, multixenobiotic resistance, and histopathology of Asian freshwater clams (\u003cem\u003eCorbicula fluminea\u003c/em\u003e). \u003cem\u003eAquatic Toxicology,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e218\u003c/strong\u003e: 105333\u003c/p\u003e\n\u003cp\u003e(17) Jeschke, P., Nauen, R., Schindler, M. and Elbert, A. (2011). Overview of the status and global strategy for neonicotinoids. \u003cem\u003eJournal of Agriculture and Food Chemistry\u003c/em\u003e \u003cstrong\u003e59\u003c/strong\u003e: 2897-2908.\u003c/p\u003e\n\u003cp\u003e(18) Goulson, D. (2013). Review: an overview of the environmental risks posed by neonicotinoid insecticides. \u003cem\u003eJournal of Applied Ecology\u003c/em\u003e \u003cstrong\u003e50\u003c/strong\u003e: 977-987.\u003c/p\u003e\n\u003cp\u003e(19) Jeschke, P. and Nauen, R. (2008). Neonicotinoids\u0026mdash;from zero to hero in insecticide chemistry. \u003cem\u003ePest Management Science\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e64\u003c/strong\u003e: 1084-1098.\u003c/p\u003e\n\u003cp\u003e(20) Lewis, K. A., Tzilivakis, J., Warner, D. J. and Green, A (2016). An international database for pesticide risk assessments and management.\u003cem\u003e\u0026nbsp;Human and Ecological Risk Assessment: An International Journal\u003c/em\u003e, \u003cstrong\u003e22\u003c/strong\u003e:1050-1064.\u003c/p\u003e\n\u003cp\u003e(21) Nnadi, J.U., Dimelu, I.N., Nwani, S.I., Madu, J.C., Atama, C.I., Attamah, G.N., Okwor, J.I. and Nwani, C.D., 2018. Biometric variations and oxidative stress responses in juvenile Clarias gariepinus exposed to Termex\u0026reg;. \u003cem\u003eAfrican Journal of Aquatic Science\u003c/em\u003e, \u003cem\u003e43\u003c/em\u003e(1), pp.27-34.\u003c/p\u003e\n\u003cp\u003e(22) Islam, M. A., Hossen, M. S., Sumon, K. A. and Rahman, M. M. (2019). Acute toxicity of imidacloprid on the developmental stages of Common Carp \u003cem\u003eCyprinus carpio\u003c/em\u003e. \u003cem\u003eToxicology and Environmental Health Sciences\u003c/em\u003e, \u003cstrong\u003e11\u003c/strong\u003e(3): 244-251.\u003c/p\u003e\n\u003cp\u003e(23) Qadir, S. and Iqbal, F. (2016). Effect of subleathal concentrtion of imidacloprid on the histology of heart, liver and kidney in \u003cem\u003eLabeo rohita\u003c/em\u003e. \u003cem\u003ePakistan journal of pharmaceutical sciences,\u003c/em\u003e \u003cstrong\u003e29\u003c/strong\u003e:2033-2038.\u003c/p\u003e\n\u003cp\u003e(24) Ogueji, E. O., Nwani, C. D., Iheanacho, S. C., Mbah, C. E., Okeke, O. C. and Yaji, A. (2018). Acute toxicity effects of ibuprofen on behaviour and haematological parameters of African catfish \u003cem\u003eClarias gariepinus\u003c/em\u003e (Burchell, 1822). \u003cem\u003eAfrican Journal of Aquatic Science,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e43\u003c/strong\u003e(2): 293\u0026ndash;303.\u003c/p\u003e\n\u003cp\u003e(25) OECD (2019). Test guideline No. 203: Fish, acute toxicity test. \u003cem\u003eOECD Guidelines for the Testing of Chemicals, Section 2.\u0026nbsp;\u003c/em\u003eOECD Publishing, Paris. doi:10.1787/9789264069961-en\u003c/p\u003e\n\u003cp\u003e(26) Reitman, S. and Frankel, S. (1957). A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. \u003cem\u003eAmerican journal of clinical pathology,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e28\u003c/strong\u003e(1): 56-63.\u003c/p\u003e\n\u003cp\u003e(27) Jendrassik, L. and Grof, P. (1938). Estimation of total serum bilirubin level by spectrophotometrically in serum and plasma. \u003cem\u003eBiochem Zeitschrift\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e297\u003c/strong\u003e: 81-89.\u003c/p\u003e\n\u003cp\u003e(28) Bern, F. P. (1955). Amylase a and b. In Methods of enzymology (Ed. Clockwi and Kalpin). Academic Press New York, USA. pp. 149-158.\u003c/p\u003e\n\u003cp\u003e(29) Bier, M. (1955). [106] Lipases: RCOOR\u0026prime;+ H2O\u0026rarr; RCOOH+ R\u0026prime; OH.\u003c/p\u003e\n\u003cp\u003e(30) Habig, W.H. and W.B. Jakoby (1981). Assays for differentiation of glutathione-S-transferases. \u003cem\u003eMethods in Enzymology,\u003c/em\u003e \u003cstrong\u003e77\u003c/strong\u003e: 398-405.\u003c/p\u003e\n\u003cp\u003e(31) Minamide, M., Hosoi, I. and Yanagi, S. (2000). CA19.9-producing testicular tumor: a case report, \u003cem\u003eHinyokika Kiyo\u003c/em\u003e, \u003cstrong\u003e46\u003c/strong\u003e(1): 45-47.\u003c/p\u003e\n\u003cp\u003e(32) Lowry, O. H., Rosenbrough, N. J., Farr, A. L. and Randall, R. J. (1951).\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003eProtein measurement with the folin phenol reagent. \u003cem\u003eJournal of Biological Chemistry,\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e193\u003c/strong\u003e: 265-275.\u003c/p\u003e\n\u003cp\u003e(33) Teimouri, M., Yeganeh, S. \u0026nbsp;Mianji, G.R., Najafi, M. and Mahjoub, S. (2019). The effect of Spirulina platensis meal on antioxidant gene expression, total antioxidant capacity, and lipid peroxidization of rainbow trout (\u003cem\u003eOncorhynchus mykiss\u003c/em\u003e). \u003cem\u003eFish Physiology and Biochemistry\u003c/em\u003e, \u003cstrong\u003e45\u003c/strong\u003e(3): 977-986.\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e(34) Beers, R. F. and Sizer, I. W. (1952). A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. \u003cem\u003eJournal of Biological Chemistry,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e195\u003c/strong\u003e(1): 133-140.\u003c/p\u003e\n\u003cp\u003e(35) Hafeman, D.G., Sunde, R.A. and Hoekstra, W.G. (1974). Effect of dietary selenium on erythrocyte and liver glutathione peroxidase in the rat. \u003cem\u003eThe Journal of nutrition\u003c/em\u003e, \u003cstrong\u003e104\u003c/strong\u003e(5): 580-587.\u003c/p\u003e\n\u003cp\u003e(36) McCord, J. M. and Fridovich, I. (1969). Superoxide dismutase: an enzymic function for erythrocuprein (hemocuprein). \u003cem\u003eJournal of Biological Chemistry,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e244\u003c/strong\u003e(22): 6049-6055.\u003c/p\u003e\n\u003cp\u003e(37) Wang, Y., Yang, G., Dai, D., Xu, Z., Cai, L., Wang, Q. and Yu, Y. (2017). Individual and mixture effects of five agricultural pesticides on zebrafish (\u003cem\u003eDanio rerio\u003c/em\u003e) larvae. \u003cem\u003eEnvironmental Science and Pollution Research\u003c/em\u003e, \u003cstrong\u003e24\u003c/strong\u003e(5): 4528-4536.\u003c/p\u003e\n\u003cp\u003e(38) Xia, X., Xia, X., Huo, W., Dong, H., Zhang, L. and Chang, Z. (2016). Toxic effects of imidacloprid on adult loach (\u003cem\u003eMisgurnus anguillicaudatus\u003c/em\u003e). \u003cem\u003eEnvironmental Toxicology and Pharmacology\u003c/em\u003e, \u003cstrong\u003e45\u003c/strong\u003e: 132-139.\u003c/p\u003e\n\u003cp\u003e(39) Naiel, M.A., Shehata, A.M., Negm, S.S., Abd El‐Hack, M.E., Amer, M.S., Khafaga, A.F., Bin‐Jumah, M. and Allam, A.A. (2020). The new aspects of using some safe feed additives on alleviated imidacloprid toxicity in farmed fish: A review. \u003cem\u003eReviews in aquaculture\u003c/em\u003e, \u003cstrong\u003e12\u003c/strong\u003e(4): 2250-2267.\u003c/p\u003e\n\u003cp\u003e(40) Qadir, S., Latif, A., Ali, M. and Iqbal, F. (2014). Effects of Imidacloprid on the hematological and serum biochemical profile of \u003cem\u003eLabeo rohita. Pakistan Journal of Zoology,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e46\u003c/strong\u003e(4): 1085-1090.\u003c/p\u003e\n\u003cp\u003e(41) Topal, A., Alak, G., Ozkaraca, M., Yeltekin, A.C., Comaklı, S., Acıl, G., Kokturk, M. and Atamanalp, M., 2017. Neurotoxic responses in brain tissues of rainbow trout exposed to imidacloprid pesticide: Assessment of 8-hydroxy-2-deoxyguanosine activity, oxidative stress and acetylcholinesterase activity. \u003cem\u003eChemosphere\u003c/em\u003e, \u003cstrong\u003e175\u003c/strong\u003e: 186-191.\u003c/p\u003e\n\u003cp\u003e(42) Rahman, A.N.A., Mansour, D.A., Abd El-Rahman, G.I., Elseddawy, N.M., Zaglool, A.W., Khamis, T., Mahmoud, S.F. and Mahboub, H.H. (2022). Imidacloprid toxicity in \u003cem\u003eClarias gariepinus\u003c/em\u003e: Protective role of dietary \u003cem\u003eHyphaene thebaica\u003c/em\u003e against biochemical and histopathological disruption, oxidative stress, immune genes expressions, and \u003cem\u003eAeromonas sobria\u003c/em\u003e infection. \u003cem\u003eAquaculture\u003c/em\u003e, \u003cstrong\u003e555\u003c/strong\u003e: 738170.\u003c/p\u003e\n\u003cp\u003e(43) Farah, A. (1991). Effect of Heavy Metal Toxicants on Liver Function and Hepatic Drug Metabolizing Enzyme System in Rabbit (Doctoral dissertation, University of the Punjab, Lahore).\u003c/p\u003e\n\u003cp\u003e(44) Ali, S.S., Qureshi, M.A., Iqbal, M.J. And Shakoori, A.R. (1992). Zinc-induced biochemical alterations in the liver of common carp, \u003cem\u003eCyprinus carpio. Punjab University Journal of Zoology,\u003c/em\u003e \u003cstrong\u003e7\u003c/strong\u003e:1-7.\u003c/p\u003e\n\u003cp\u003e(45) Yousafzai, A. M. and Shakoori, A. R. (2011). Hepatic responses of a freshwater fish against aquatic pollution. \u003cem\u003ePakistan Journal of Zoology,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e43\u003c/strong\u003e(2): 209-221.\u003c/p\u003e\n\u003cp\u003e(46) Desai, B. and Parikh, P. (2013). Biochemical alterations on exposure of imidacloprid and curzate on fresh water fish \u003cem\u003eOreochromis mossambicus\u003c/em\u003e and \u003cem\u003eLabeo rohita\u003c/em\u003e. \u003cem\u003eIndian Journal of Forensic Medicine \u0026amp; Toxicology\u003c/em\u003e, \u003cstrong\u003e7\u003c/strong\u003e(2): 87-91.\u003c/p\u003e\n\u003cp\u003e(47) El Euony, O.I., Elblehi, S.S., Abdel-Latif, H.M., Abdel-Daim, M.M. and El-Sayed, Y.S., 2020. Modulatory role of dietary \u003cem\u003eThymus vulgaris\u003c/em\u003e essential oil and \u003cem\u003eBacillus subtilis\u003c/em\u003e against thiamethoxam-induced hepatorenal damage, oxidative stress, and immunotoxicity in African catfish (\u003cem\u003eClarias garipenus\u003c/em\u003e). \u003cem\u003eEnvironmental Science and Pollution Research\u003c/em\u003e, \u003cstrong\u003e27\u003c/strong\u003e(18): 23108-23128.\u003c/p\u003e\n\u003cp\u003e(48) Rao, V. J. (2006). Sublethal effects of an organophosphorus insecticide (RPR-II) on biochemical parameters of Tilapia, \u003cem\u003eOreochromis mossambicus\u003c/em\u003e. \u003cem\u003eComparative Biochemistry and Physiology Part C: Toxicology and Pharmacology,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e143\u003c/strong\u003e(4): 492-498.\u003c/p\u003e\n\u003cp\u003e(49) Sahu, S., Debasmita, S.and Mohapatra, A. K. (2016). Impact of cadmium on some biochemical profiles of the fresh water cat fish \u003cem\u003eClarias gariepinus\u003c/em\u003e (burchell, 1822). \u003cem\u003eThe Ecoscan,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e9\u003c/strong\u003e: 257-265.\u003c/p\u003e\n\u003cp\u003e(50) Sanna, S.G., Tabassum, S., Noreen, S., Mahmood, S., Rehman, M.U., Ahmad, B., Kabir, M., Sajid, M. and Khan, M.F. (2021). Biochemical, Endocrine and Genetic Impairments in Response to Agrochemicals Intoxication in Common Carp (Cyprinus carpio). International Journal of Agriculture \u0026amp; Biology. doi: 10.17957/IJAB/15.1787.\u003c/p\u003e\n\u003cp\u003e(51) Samanta, P., Pal, S., Mukherjee, A.K., Senapati, T. and Ghosh, A.R. (2019). March. Effects of Glyphosate-Based Herbicide (Excel Mera 71) on Digestive Enzymes and Recovery Pattern in Teleostean Fishes. \u003cem\u003eProceedings of the Zoological Society,\u003c/em\u003e \u003cstrong\u003e72\u003c/strong\u003e(1): 54-60.\u003c/p\u003e\n\u003cp\u003e(52) Samanta, P., Pal, S., Mukherjee, A.K., Senapati, T., Kole, D. and Ghosh, A.R. (2014). Effects of almix herbicide on profile of digestive enzymes of three freshwater teleostean fishes in rice field condition. \u003cem\u003eToxicology Reports\u003c/em\u003e, \u003cstrong\u003e1\u003c/strong\u003e: 379-384.\u003c/p\u003e\n\u003cp\u003e(53) Samanta, P., Pal, S., Mukherjee, A. K., Senapati, T. and Ghosh, A. R. (2016). Alterations in digestive enzymes of three freshwater teleostean fishes by almix herbicide: A comparative study. \u003cem\u003eProceedings of the Zoological Society,\u003c/em\u003e \u003cstrong\u003e69\u003c/strong\u003e(1): 61-66.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e(54) Cornelius, C. E. (1991). Bile Pigments in Fishes: A Review\u003cem\u003e. Veterinary Clinical Pathology,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e20\u003c/strong\u003e(4):\u003cem\u003e\u0026nbsp;\u003c/em\u003e106\u0026ndash;116\u003cem\u003e.\u003c/em\u003e\u003c/p\u003e\n\u003cp\u003e(55) Ujowundu, C. O., Nnanna, C. G., Ndubuisi, E. U., Ngwu, P. C., Uzoma, C. W. and Ezeji, E. U. (2017). Glyphosate-based pesticide-induced biochemical changes in hepatic and renal tissues of \u003cem\u003eClarias gariepinus\u003c/em\u003e. \u003cem\u003eFuto Journal Series,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e3\u003c/strong\u003e(1): 225-235.\u003c/p\u003e\n\u003cp\u003e(56) Inyang, I. R., Akparanta, O. J. and Izah, S. C. (2019). Evaluation of some blood cells and metabolites of \u003cem\u003eHeterobranchus bidorsalis\u003c/em\u003e (hybrid) exposed to sublethal concentrations of sarosate. \u003cem\u003eInternational Journal of Research in Environmental Science,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e5\u003c/strong\u003e(1): 17-23.\u003c/p\u003e\n\u003cp\u003e(57) Vitek, L. and Ostrou, J. D. (2009). Bilirubin chemistry and metabolism, harmful and protective aspects. \u003cem\u003eCurrent Pharmaceutical Design,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e15\u003c/strong\u003e: 2869-2883.\u003c/p\u003e\n\u003cp\u003e(58) Kumar, V. A., Janaiah, C. and Venkateshwarlu, P. (2010). Effect of thiamethoxam alters serum biochemical parameters in \u003cem\u003eChanna punctatus\u003c/em\u003e (Bloch). \u003cem\u003eAsian Journal of Bio Science,\u003c/em\u003e \u003cstrong\u003e5\u003c/strong\u003e(1): 106-110.\u003c/p\u003e\n\u003cp\u003e(59) Nagahama, Y. and Yamashita, M. (2008). Regulation of oocyte maturation in fish. \u003cem\u003eDevelopment, Growth \u0026amp; Differentiation\u003c/em\u003e, \u003cstrong\u003e50\u003c/strong\u003e: S195\u0026ndash;S219.\u003c/p\u003e\n\u003cp\u003e(60) Azcoitia, I., Barreto, G.E. and Garcia-Segura, L.M. (2019). Molecular mechanisms and cellular events involved in the neuroprotective actions of estradiol. Analysis of sex differences. \u003cem\u003eFrontiers in Neuroendocrinology\u003c/em\u003e, \u003cstrong\u003e55\u003c/strong\u003e: 100787.\u003c/p\u003e\n\u003cp\u003e(61) Manolagas, S. C. and Kousteni, S. (2001). Perspective: Nonreproductive sites of action of reproductive hormones. \u003cem\u003eEndocrinology\u003c/em\u003e, \u003cstrong\u003e142\u003c/strong\u003e(6): 2200\u0026ndash;2204.\u003c/p\u003e\n\u003cp\u003e(62) Bouman, A., Heineman, M. J. and Faas, M. M. (2005). Sex hormones and the immune response in humans. \u003cem\u003eHuman Reproduction Update\u003c/em\u003e, \u003cstrong\u003e111\u003c/strong\u003e(4): 411\u0026ndash;423.\u003c/p\u003e\n\u003cp\u003e(63)\u0026nbsp;Jin, J., Kurobe, T., Ram\u0026iacute;rez-Duarte, W. F., Bolotaolo, M. B., Lam, C. H., Pandey, P. K., \u0026hellip; Teh, S. J. (2018). Sub-lethal effects of herbicides penoxsulam, imazamox, fluridone and glyphosate on Delta Smelt\u003cem\u003e\u0026nbsp;\u003c/em\u003e(\u003cem\u003eHypomesus transpacificus\u003c/em\u003e).\u003cem\u003e\u0026nbsp;Aquatic Toxicology,\u0026nbsp;\u003c/em\u003e\u003cstrong\u003e197\u003c/strong\u003e: 79\u0026ndash;88.\u003c/p\u003e\n\u003cp\u003e(64)\u0026nbsp;Tripathy, P.S., Parhi, J. and Mandal, S.C., 2021. Steroids and Its Receptors in Fish Reproduction. In \u003cem\u003eRecent updates in molecular Endocrinology and Reproductive Physiology of Fish,\u003c/em\u003e (pp. 53-61). Springer, Singapore.\u003c/p\u003e\n\u003cp\u003e(65) Sanderson, C. J. (1992). Interleukin-5, Eosinophils, and Disease. \u003cem\u003eBlood\u003c/em\u003e, \u003cstrong\u003e79\u003c/strong\u003e(12): 3101\u0026ndash;3109.\u003c/p\u003e\n\u003cp\u003e(66) Chaves-Pozo, E., Garc\u0026iacute;a-Ayala, A. and Cabas, I. (2018). Effects of sex steroids on fish leukocytes. \u003cem\u003eBiology\u003c/em\u003e, \u003cstrong\u003e7\u003c/strong\u003e: 9.\u003c/p\u003e\n\u003cp\u003e(67) Chakrabarty, S., Rajakumar, A., Raqhuveer, K., Sridevi, P., Mohanachary, A. and Prathibha Y. (2012). Endosulfan and flutamide, alone and in combination, target ovarian growth in juvenile catfish, \u003cem\u003eClarias batrachus\u003c/em\u003e. \u003cem\u003eComparative Biochemistry and Physiology,\u003c/em\u003e \u003cstrong\u003e155\u003c/strong\u003e: 491\u0026ndash;497.\u003c/p\u003e\n\u003cp\u003e(68) Agbohessi, T.P., Toko, I.I., Atchou, V., Tonato, R., Mandiki, S.N.M. and Kestemont, P. (2015). Pesticides used in cotton production affect reproductive development, endocrine regulation, liver status and offspring fitness in African catfish \u003cem\u003eClarias gariepinus\u003c/em\u003e (Burchell, 1822). \u003cem\u003eComparative Biochemistry and Physiology,\u003c/em\u003e \u003cstrong\u003e167\u003c/strong\u003e: 157-172\u003c/p\u003e\n\u003cp\u003e(69) Anjali, V.R., Remya, V.S., Reshmi, S., Mahim, S.S. and Devi, C.A. (2019). Impact of Bisphenol S as an Endocrine Disruptor in a Freshwater Fish, \u003cem\u003eOreochromis mossambicus\u003c/em\u003e. \u003cem\u003eJournal of Endocrinology and Reproduction\u003c/em\u003e, \u003cstrong\u003e23\u003c/strong\u003e(2): 49-63.\u003c/p\u003e\n\u003cp\u003e(70) Marlatt, V.L., Leung, T.Y.G., Calbick, S., Metcalfe, C. and Kennedy, C. (2019). Sub-lethal effects of a neonicotinoid, clothianidin, on wild early life stage sockeye salmon (O\u003cem\u003encorhynchus nerka\u003c/em\u003e). \u003cem\u003eAquatic Toxicology\u003c/em\u003e, \u003cstrong\u003e217\u003c/strong\u003e: 105335.\u003c/p\u003e\n\u003cp\u003e(71) Blanton, M.L. and Specker, J.L. (2007). The hypothalamic-pituitary-thyroid (HPT) axis in fish and its role in fish development and reproduction. \u003cem\u003eCritical Reviews in Toxicology\u003c/em\u003e, \u003cstrong\u003e37\u003c/strong\u003e(1-2): 97-115.\u003c/p\u003e\n\u003cp\u003e(72) Lacasa\u0026ntilde;a, M., L\u0026oacute;pez-Flores, I., Rodr\u0026iacute;guez-Barranco, M., Aguilar-Gardu\u0026ntilde;o, C., Blanco-Mu\u0026ntilde;oz, J., P\u0026eacute;rez-M\u0026eacute;ndez, O., Gamboa, R., Bassol, S. and Cebrian, M.E. (2010). Association between organophosphate pesticides exposure and thyroid hormones in floriculture workers. \u003cem\u003eToxicology and Applied Pharmacology\u003c/em\u003e, \u003cstrong\u003e243\u003c/strong\u003e(1): 19-26.\u003c/p\u003e\n\u003cp\u003e(73) Yu, L., Han, Z. and Liu, C. (2015). A review on the effects of PBDEs on thyroid and reproduction systems in fish. \u003cem\u003eGeneral and Comparative Endocrinology\u003c/em\u003e, \u003cstrong\u003e219\u003c/strong\u003e: 64-73.\u003c/p\u003e\n\u003cp\u003e(74) Campinho, M.A. (2019). Teleost metamorphosis: the role of thyroid hormone. \u003cem\u003eFrontiers in Endocrinology\u003c/em\u003e, \u003cstrong\u003e10\u003c/strong\u003e: 383.\u003c/p\u003e\n\u003cp\u003e(75) Van der Geyten, S., Mol, K.A., Pluymers, W., K\u0026uuml;hn, E.R. and Darras, V.M. (1998). Changes in plasma T3 during fasting/refeeding in tilapia (\u003cem\u003eOreochromis niloticus\u003c/em\u003e) are mainly regulated through changes in hepatic type II iodothyronine deiodinase. \u003cem\u003eFish Physiology and Biochemistry\u003c/em\u003e, \u003cstrong\u003e19\u003c/strong\u003e(2): 135-143.\u003c/p\u003e\n\u003cp\u003e(76) Power, D.M., Llewellyn, L., Faustino, M., Nowell, M.A., Bj\u0026ouml;rnsson, B.T., Einarsd\u0026oacute;ttir, I.E., Canario, A.V. and Sweeney, G.E. (2001). Thyroid hormones in growth and development of fish. \u003cem\u003eComparative Biochemistry and Physiology Part C: Toxicology \u0026amp; Pharmacology\u003c/em\u003e, \u003cstrong\u003e130\u003c/strong\u003e(4): 447-459.\u003c/p\u003e\n\u003cp\u003e(77) Ghayyur, S., Khan, M.F., Tabassum, S., Ahmad, M.S., Sajid, M., Badshah, K., Khan, M.A., Ghayyur, S., Khan, N.A., Ahmad, B. and Qamer, S. (2021). A comparative study on the effects of selected pesticides on hemato-biochemistry and tissue histology of freshwater fish \u003cem\u003eCirrhinus mrigala\u0026nbsp;\u003c/em\u003e(Hamilton, 1822). \u003cem\u003eSaudi Journal of Biological Sciences\u003c/em\u003e, \u003cstrong\u003e28\u003c/strong\u003e(1): 603-611.\u003c/p\u003e\n\u003cp\u003e(78) Saha, S., Chukwuka, A.V., Mukherjee, D., Patnaik, L., Nayak, S., Dhara, K., Saha, N.C. and Faggio, C., 2021. Chronic effects of Diazinon\u0026reg; exposures using integrated biomarker responses in freshwater walking catfish, Clarias batrachus. \u003cem\u003eApplied Sciences\u003c/em\u003e, \u003cstrong\u003e11\u003c/strong\u003e(22): 10902.\u003c/p\u003e\n\u003cp\u003e(79) Yu, L.Q., Zhao, G.F., Feng, M., Wen, W., Li, K., Zhang, P.W., Peng, X., Huo, W.J. and Zhou, H.D. (2014). Chronic exposure to pentachlorophenol alters thyroid hormones and thyroid hormone pathway mRNAs in zebrafish. \u003cem\u003eEnvironmental Toxicology and Chemistry\u003c/em\u003e, \u003cstrong\u003e33\u003c/strong\u003e(1): 170-176.\u003c/p\u003e\n\u003cp\u003e(80) Ndonwi, E.N., Atogho-Tiedeu, B., Lontchi-Yimagou, E., Shinkafi, T.S., Nanfa, D., Balti, E.V., Indusmita, R., Mahmood, A., Katte, J.C., Mbanya, A. and Matsha, T. (2019). Gestational exposure to pesticides induces oxidative stress and lipid peroxidation in offspring that persist at adult age in an animal model. \u003cem\u003eToxicological Research\u003c/em\u003e, \u003cstrong\u003e35\u003c/strong\u003e(3): 241-248.\u003c/p\u003e\n\u003cp\u003e(81) Abdel-Rahman Mohamed, A., Abdel Rahman, A.N., Salem, G.A., Deib, M.M.E., Nassan, M.A., Rhouma, N.R. and Khater, S.I. (2021). The antioxidant role of a taurine-enriched diet in combating the immunotoxic and inflammatory effects of pyrethroids and/or carbamates in \u003cem\u003eOreochromis niloticus\u003c/em\u003e. \u003cem\u003eAnimals\u003c/em\u003e, \u003cstrong\u003e11\u003c/strong\u003e(5): 1318.\u003c/p\u003e\n\u003cp\u003e(82) Borgesa, A.C.P., Piassao, J.F.G., Albani, S.M., Albertoni, E.F., Martins, M.C., Cansian, R.L., Valduga, A.T., Hepp, L.U. and Mielniczki-Pereira, A.A. (2022). Multiple metals and agricultural use affects oxidative stress biomarkers in freshwater Aegla crabs/Presenca de multiplos metais e agricultura afetam biomarcadores de estresse oxidativo em caranguejos de agua doce (Aegla). \u003cem\u003eBrazilian Journal of Biology\u003c/em\u003e, 82: e230147.\u003c/p\u003e\n\u003cp\u003e(83) Hellou, J., Ross, N.W. and Moon, T.W. (2012). Glutathione, glutathione S-transferase, and glutathione conjugates, complementary markers of oxidative stress in aquatic biota. \u003cem\u003eEnvironmental Science and Pollution Research\u003c/em\u003e, \u003cstrong\u003e19\u003c/strong\u003e(6): 2007-2023.\u003c/p\u003e\n\u003cp\u003e(84) Ge, W., Yan, S., Wang, J., Zhu, L., Chen, A. and Wang, J. (2015). Oxidative stress and DNA damage induced by imidacloprid in zebrafish (\u003cem\u003eDanio rerio\u003c/em\u003e). \u003cem\u003eJournal of agricultural and food chemistry\u003c/em\u003e, \u003cstrong\u003e63\u003c/strong\u003e(6): 1856-1862.\u003c/p\u003e\n\u003cp\u003e(85) John, S., Kale, M., Rathore, N. and Bhatnagar, D. (2001). Protective effect of vitamin E in dimethoate and malathion induced oxidative stress in rat erythrocytes. \u003cem\u003eThe Journal of Nutritional Biochemistry\u003c/em\u003e, \u003cstrong\u003e12\u003c/strong\u003e(9): 500-504.\u003c/p\u003e\n\u003cp\u003e(86) Svensson, R., Rinaldi, R., Swedmark, S. and Morgenstern, R. (2000). Reactivity of cysteine-49 and its influence on the activation of microsomal glutathione transferase 1: evidence for subunit interaction. \u003cem\u003eBiochemistry\u003c/em\u003e, \u003cstrong\u003e39\u003c/strong\u003e(49): 15144-15149.\u003c/p\u003e\n\u003cp\u003e(87) Chang, Y., Mao, L., Zhang, L., Zhang, Y. and Jiang, H. (2020). Combined toxicity of imidacloprid, acetochlor, and tebuconazole to zebrafish (\u003cem\u003eDanio rerio\u003c/em\u003e): acute toxicity and hepatotoxicity assessment. \u003cem\u003eEnvironmental Science and Pollution Research\u003c/em\u003e, \u003cstrong\u003e27\u003c/strong\u003e(10): 10286-10295.\u003c/p\u003e\n\u003cp\u003e(88) G\u0026uuml;nal, A.\u0026Ccedil;., Erkmen, B., Pa\u0026ccedil;al, E., Arslan, P., Yildirim, Z. and Erko\u0026ccedil;, F. (2020). Sub-lethal Effects of Imidacloprid on Nile tilapia (\u003cem\u003eOreochromis niloticus\u003c/em\u003e). \u003cem\u003eWater, Air, \u0026amp; Soil Pollution\u003c/em\u003e, \u003cstrong\u003e231\u003c/strong\u003e(1): 1-10.\u003c/p\u003e\n\u003cp\u003e(89) Safi, F., Beger, H.G., Bittner, R., B\u0026uuml;chler, M. and Krautzberger, W. (1986). CA 19‐9 and pancreatic adenocarcinoma. \u003cem\u003eCancer\u003c/em\u003e, \u003cstrong\u003e57\u003c/strong\u003e(4): 779-783.\u003c/p\u003e\n\u003cp\u003e(90) Wahab, M.A., El-Bendary, M., Ali, M.A., Shehta, A., Abed, S., Shehatta, A.S., Salah, M. and El-Gilany, A.H. (2018). Serum Levels of Heavy Metals in Cholangiocarcinoma Patients from the Nile Delta Region of Egypt: A Single Centre Study. \u003cem\u003eSurg. Gastroenterol\u003c/em\u003e, \u003cstrong\u003e23\u003c/strong\u003e(3): 186-190.\u003c/p\u003e\n\u003cp\u003e(91) Abdel-Hamid, O.M., Nafee, A.A., MA, E. and MA, E. (2018). The ameliorative effect of Vitamin C in experimentally induced colon cancer in rats. \u003cem\u003eBenha Veterinary Medical Journal\u003c/em\u003e, \u003cstrong\u003e34\u003c/strong\u003e(1): 329-343.\u003c/p\u003e\n\u003cp\u003e(92) Kim, S., Park, B.K., Seo, J.H., Choi, J., Choi, J.W., Lee, C.K., Chung, J.B., Park, Y. and Kim, D.W. (2020). Carbohydrate antigen 19-9 elevation without evidence of malignant or pancreatobiliary diseases. \u003cem\u003eScientific reports\u003c/em\u003e, \u003cstrong\u003e10\u003c/strong\u003e: 8820.\u003c/p\u003e\n\u003cp\u003e(93) Sepici-Din\u0026ccedil;el, A., Benli, A.\u0026Ccedil;.K., Selvi, M., Sarıkaya, R., Şahin, D., \u0026Ouml;zkul, I.A. and Erko\u0026ccedil;, F. (2009). Sublethal cyfluthrin toxicity to carp (\u003cem\u003eCyprinus carpio\u003c/em\u003e L.) fingerlings: biochemical, hematological, histopathological alterations. \u003cem\u003eEcotoxicology and Environmental Safety\u003c/em\u003e, \u003cstrong\u003e72\u003c/strong\u003e(5): 1433-1439.\u003c/p\u003e\n\u003cp\u003e(94) Nemeth, E., Baird, A.W. and O\u0026rsquo;Farrelly, C., 2009, September. Microanatomy of the liver immune system. \u003cem\u003eSeminars in immunopathology,\u003c/em\u003e \u003cstrong\u003e31\u003c/strong\u003e(3): 333-343.\u003c/p\u003e\n\u003cp\u003e(95) Rout, P.C. and Naik, B.N., 1998. Elevation of blood histamine and C-RP durings short-term lead exposure in \u003cem\u003eClarias batrachus\u003c/em\u003e (Linn.). \u003cem\u003eJournal of Nature Conservators\u003c/em\u003e, \u003cstrong\u003e10\u003c/strong\u003e(2): 169-173.\u003c/p\u003e\n\u003cp\u003e(96) Chatterjee, R. (2016). Estimation of endosulfan toxicity on blood biochemistry of channel catfish Clarias batrachus Linn. \u003cem\u003eSouth Indian Journal of Biological Sciences\u003c/em\u003e, \u003cstrong\u003e2\u003c/strong\u003e(4): 445-450.\u003c/p\u003e\n\u003cp\u003e(97) Paul, I., Mandal, C. and Mandal, C. (1998). Effect of environmental pollutants on the C-reactive protein of a freshwater major carp, \u003cem\u003eCatla catla\u003c/em\u003e. \u003cem\u003eDevelopmental \u0026amp; Comparative Immunology\u003c/em\u003e, \u003cstrong\u003e22\u003c/strong\u003e(5-6): 519-532.\u003c/p\u003e\n\u003cp\u003e(98) Kodama, H., Matsuoka, Y., Tanaka, Y., Liu, Y., Iwasaki, T. and Watarai, S. (2004). Changes of C-reactive protein levels in rainbow trout (\u003cem\u003eOncorhynchus mykiss\u003c/em\u003e) sera after exposure to anti-ectoparasitic chemicals used in aquaculture. \u003cem\u003eFish \u0026amp; shellfish immunology\u003c/em\u003e, \u003cstrong\u003e16\u003c/strong\u003e(5): 589-597.\u003c/p\u003e\n\u003cp\u003e(99) Tarnawska, M., Augustyniak, M., Łaszczyca, P., Migula, P., Irnazarow, I., Krzyżowski, M. and Babczyńska, A. (2019). Immune response of juvenile common carp (\u003cem\u003eCyprinus carpio\u003c/em\u003e L.) exposed to a mixture of sewage chemicals. \u003cem\u003eFish and Shellfish Immunology\u003c/em\u003e, \u003cstrong\u003e88\u003c/strong\u003e: 17-27.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"University of Benin","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"behavioral markers, imidacloprid, oxidative stress, thyroid hormones, reproductive hormones, hepatic markers, tumor marker, inflammatory marker ","lastPublishedDoi":"10.21203/rs.3.rs-2193834/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2193834/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eImidacloprid is among the most widely used insecticides and may contaminate surface waters, yet there is limited information available on their effects on fish. In the present study, juvenile catfish (\u003cem\u003eClarias gariepinus\u003c/em\u003e) were exposed to 100, 130, 160, and 190 mg/L of imidacloprid for 96 hours, and the effects on mortality, behavior, hepatic and endocrine functions, oxidative stress, and tumor and inflammatory responses were investigated. The 96 hours LC\u003csub\u003e50\u003c/sub\u003e was 166.60 (143.38-193.59 95% confidence intervals) mg/L with mortality recorded at concentrations\u0026thinsp;\u0026ge;\u0026thinsp;130 mg/L. Exposure to imidacloprid induced behavioral alterations and clinical symptoms including gulping, hypoactivity, abnormal surface distribution, loss of buoyancy, and excessive mucus secretion. Furthermore, amylase, lipase, conjugate bilirubin, carbohydrate antigen 19\u0026thinsp;\u0026minus;\u0026thinsp;9 (CA 19\u0026thinsp;\u0026minus;\u0026thinsp;9), thyroid stimulating hormone (TSH), and glutathione peroxidase (GPx) activity were significantly increased, while significant reductions were observed in triiodothyronine (T3) and superoxide dismutase (SOD) activity. Non-significant changes were observed in aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin, 17β- estradiol (E2), thyroxine (T4), C-reactive protein (CRP), glutathione S-transferases (GST), total antioxidant capacity (TAC) and catalase (CAT) activity. Our results demonstrate that imidacloprid causes behavioral, hepatic, and endocrine toxicity in catfish as well as oxidative stress and tumor marker elevation. Our findings show that short-term exposure to imidacloprid triggers biochemical and physiological responses in juvenile catfish.\u003c/p\u003e","manuscriptTitle":"Does short-term exposure to a Neonicotinoid insecticide trigger Biochemical and Physiological responses in Juvenile catfish?","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-10-24 18:08:53","doi":"10.21203/rs.3.rs-2193834/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"9eab6ae2-bdd3-4c20-a451-df1d3fe4c4b8","owner":[],"postedDate":"October 24th, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":16440288,"name":"Toxicology"},{"id":16440289,"name":"Behavioral Ecology"},{"id":16440290,"name":"Animal Physiology"}],"tags":[],"updatedAt":"2022-10-24T18:08:53+00:00","versionOfRecord":[],"versionCreatedAt":"2022-10-24 18:08:53","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-2193834","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2193834","identity":"rs-2193834","version":["v1"]},"buildId":"GqpaHPwrfC8PjnIFayRh5","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}