Nigella sativa seeds mitigate the hepatic histo-architectural and ultrastructural changes induced by 4-nonylphenol in Clarias gariepinus.

Due to its prevalence in aquatic environments and potential cytotoxicity, 4-nonylphenol (4-NP) has garnered considerable attention. As a medicinal plant with numerous biological activities, Nigella sativa (black seed or black cumin) seed (NSS) is widely utilized throughout the world. Consequently, this study aimed to examine the potential protective effects of NSS against 4-NP-induced hepatotoxicity in African catfish (Clarias gariepinus). To achieve this objective, 18 fish (351 ± 3 g) were randomly divided into three equal groups for 21 days. The first group serves as a control which did not receive any treatment except the basal diet. The second and third groups were exposed to 4-NP at a dose of 0.1 mg L-1 of aquarium water and fed a basal diet only or supplemented with 2.5% NSS, respectively. The histological, histochemical, and ultrastructural features of the liver were subsequently evaluated as a damage biomarker of the hepatic tissue. Our results confirmed that 4-NP was a potent hepatotoxic agent, as 4-NP-intoxicated fish exhibited many lesions. Steatohepatitis, ballooning degeneration, sclerosing cholangitis, and coagulative necrosis of melanomacrophagecenters (MMCs) were observed. Hemosiderin, lipofuscin pigments, and proliferation of fibroblasts, kupffer cells, and telocytes were also demonstrated in the livers of 4-NP-intoxicated fish. In addition, decreased glycogen content and increased collagen deposition were observed in the hepatic tissue. Hepatocytes exhibited ultrastructural alterations in the chromatin, rough endoplasmic reticulum, smooth endoplasmic reticulum, mitochondria, lysosomes, and peroxisomes. Co-administration of 2.5% NSS to 4-NP-intoxicated fish significantly reduced these hepatotoxic effects. It nearly preserved the histological, histochemical, and ultrastructural integrity of hepatic tissue.

The mitigating effect of Spirulina (Arthrospira platensis) on the hemotoxicity of gibberellic acid on juvenile tilapia (Oreochromis niloticus).

The use of plant growth regulators has led to environmental contamination of water bodies that occur adjacent to agricultural areas. Some of these chemicals are bioactive, not only to plants, but also to non-target exposed biota, namely of the aquatic compartment. Previous work demonstrated the establishment of hepato- and nephrotoxic effects in juvenile tilapia (Oreochromis niloticus) exposed via aquatic media to gibberellic acid (GA3), which is among the most used plant growth regulators, in agricultural practices. Here, we investigated the effect of GA3 on hematological indices, poikilocytosis, nuclear abnormalities, and genotoxic indices measured in Nile tilapia (Oreochromis niloticus), as well as the putative protective effects of dietary supplementation of Spirulina (Arthrospira platensis). Fish were evenly assorted into 5 groups: group I served as a control, and groups II-V were fed diets supplemented with Spirulina at rates of 0 g/kg, 5 g/kg, 20 g/kg, and 100 g/kg, respectively, for 2 months before being exposed to 150 mg/L GA3. The results revealed that GA3 exposure decreased significantly all hematological indices (P < 0.05), except leucocytes and mean corpuscular hemoglobin concentration (MCHC), compared to the control group (P > 0.05). GA3 exposure increased significantly the percentage of nuclear abnormalities, altered erythrocytes and the percentages of tail DNA, compared to the control group (P < 0.05). Spirulina supplementation restored the hematological, poikilocytosis, nuclear abnormalities, and the percentages of tail DNA to near normal levels. The 100 g/kg SP treatment was the most effective in attaining such effect, showing concentration-dependency. The present study reinforces our findings of the toxicity of GA3 on O. niloticus and suggests that the addition of Spirulina to fish diet can mitigate the hemotoxic effects of GA3.

Bioremediation of hemotoxic and oxidative stress induced by polyethylene microplastic in Clarias gariepinus using lycopene, citric acid, and chlorella.

Despite extensive research on the toxic effects of microplastics (MPs), there is no obtainable data on the use of phytobioremediation against MPs toxicity in fish. This study aimed to investigate the protective role of lycopene, citric acid, and chlorella against the toxic effects of MPs in African catfish (Clarias gariepinus) using hematology, biochemical, antioxidants, erythron profiles (poikilocytosis and nuclear abnormalities) and the accumulation of MPs in tissues as biomarkers. Five groups of fish received: normal diet (control); MPs (500 mg/kg diet) (Group 2); MPs (500 mg/kg diet) + lycopene (500 mg/kg diet) (Group 3); MPs (500 mg/kg diet) + citric acid (30 g/kg diet) (Group 4); and MPs (500 mg/kg diet) + chlorella (50 g/kg diet) (Group 5) for 15 days. Group 2 had significantly higher amounts of MPs in the stomach, gills, and feces, electrolyte imbalances (HCO3, Fe, Na+, K+, Ca+2, Cl-, and anion gap, hematobiochemical alterations, and decreases in the activities of superoxide dismutase, catalase, total antioxidant capacity, and glutathione S-transferases compared to the control group. Additionally, Group 2 had significant increase in the percentage of poikilocytosis, and nuclear abnormalities in RBC's compared to the control group. The co-treatment of MPs-exposed fish with lycopene, citric acid, and chlorella-supplemented diets ameliorated the hematological, biochemical, and erythron profile alterations, but only slightly enhanced the antioxidant activity. Overall, lycopene, citric acid, and chlorella can be recommended as a feed supplement to improve hematobiochemical alterations and oxidative damage induced by MPs toxicity in the African catfish (C. gariepinus).

Impacts of microplastics on reproductive performance of male tilapia (Oreochromis niloticus) pre-fed on Amphora coffeaeformis.

This study examines the potential defending effects of the diatom, Amphora coffeaeformis, as a feed additive versus the deleterious effects (mainly on gonads) caused by microplastics (MPs) in Nile tilapia, Oreochromes niloticus. Groups of male tilapia were pre-fed diets with four different supplementation levels of A. coffeaeformis (0%, 2.5%, 5%, and 7.5%) for 70 days, then exposed to 10 mg/L MPs for 15 days. Thereafter, samples were taken from the four experimental groups and the control fish group, for evaluating blood picture, erythrocytes alterations, biochemical parameters, catalase (CAT), superoxide dismutase (SDO), and total antioxidant capacity (TAC). In addition, male reproductive performance was assessed by quantifying the follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (T), also testicular sections and GSI% were also assessed. The results showed that the number of leucocytes were significantly (p < 0.05) higher, but the number of red blood cells (RBCs), the level of Hemoglobin (Hb), the percentage of Hematocrit (Ht), the number of platelets, and the eosinophil percentages were significantly decreased (p < 0.05) when fish exposed to MPs. Biochemical parameters (ALP, glucose, uric acid, albumin, and A/G ratio) were significantly increased after MP exposure compared with the control group. Furthermore, MPs induced a significant decline in both serum LH and T levels. Testicular, histological, degenerative changes and testis-ova were found in the MP-exposed fish. Thus, A. coffeaeformis supplementation displayed ameliorative properties that detoxified the negative effects of MPs. This study provides a better understanding of the reproductive injuries caused by MPs exposure and evidence for the use of A. coffeaeformis as a natural remedy in freshwater tilapia.

Investigating the effects of copper sulfate and copper oxide nanoparticles in Nile tilapia (Oreochromis niloticus) using multiple biomarkers: the prophylactic role of Spirulina.

Copper has toxic effects in fish, whereas the cyanobacterium Spirulina reportedly has protective effects against metal toxicity in various animal species. The current study, therefore, aimed to investigate the prophylactic role of Spirulina platensis against the effects of copper sulfate (CuSO4) and copper oxide nanoparticles (CuO-NPs) in Nile tilapia (Oreochromis niloticus). Biochemical, antioxidant, erthyron profile and histopathological endpoints were assessed after for 15 days of exposure in five separate treatment groups: (1) fish pre-fed the normal diet (control), (2) fish pre-fed the normal diet and exposed to 15 mg/L of CuSO4, (3) fish pre-fed the normal diet augmented with 0.25% Spirulina and exposed to 15 mg/L of CuSO4, (4) fish pre-fed the normal diet and exposed to 15 mg/L of CuO-NPs, and (5) fish pre-fed the normal diet augmented with 0.25% Spirulina and exposed to 15 mg/L CuO-NPs. Exposure to CuSO4 or CuO-NPs significantly increased superoxide dismutase and catalase activities in fish, as well as serum total protein, glucose, aspartate aminotransferase, alanine aminotransferase, creatinine, and uric acid concentrations. In contrast, most hematological indices in fish significantly decreased after CuSO4 or CuO-NPs exposure. Moreover, CuSO4 and CuO-NPs caused a significant increase in the percentage of poikilocytosis and nuclear abnormalities of red blood cells, as well as histopathological changes in the brain, liver, intestine, and kidneys. Importantly, Spirulina supplementation mitigated against physiological disruption caused by CuSO4 or CuO-NPs.

Cytoprotective effect of Nigella sativa seed on 4-nonylphenol-induced renal damage in the African catfish (Clarias gariepinus).

4-Nonylphenol (4-NP) is a nephrotoxic substance that is highly prevalent in aquatic environments. Nigella sativa seed (NSS) has many biological activities and is widely used throughout the world as a medicinal product. Therefore, in the present study, we investigated the cytoprotective effect of NSS on 4-NP-induced renal damage in African catfish (Clarias gariepinus). Thirty fish were divided into five equal groups: an untreated control group and four groups that were challenged with 4-NP at a dose of 0.1 mg L-1 of aquarium water and fed a basal diet supplemented with 0%, 1%, 2.5%, and 5% NSS, respectively, for 3 weeks. Histological, histochemical, and ultrastructural features of the kidney were then assessed as biomarkers for renal tissue damage. Our results confirmed that 4-NP was a potent cytotoxic agent for the kidney tissue and induced renal damage, with 4-NP-intoxicated fish showing necrosis in the epithelial cells of the renal corpuscles, renal proximal convoluted tubules, and intertubular hematopoietic tissue, as well as loss of or a decrease in microvilli, a decrease in mitochondria, and an increase in the lysosomes in the epithelial cells of the proximal convoluted tubules. The kidneys of 4-NP-intoxicated fish also showed increased numbers of Perls' Prussian blue-positive melanomacrophage centers and intraepithelial T-lymphocytes in the proximal convoluted tubules and plasma cells. The administration of NSS to 4-NP-challenged fish significantly minimized the cytotoxic effect of 4-NP, maintaining the normal kidney structure, with concentrations of 2.5% and 5% of feed being most effective for protecting the kidney against 4-NP-induced renal damage.

Ameliorative effect of Spirulina platensis against lead nitrate-induced cytotoxicity and genotoxicity in catfish Clarias gariepinus.

The present study was designed to investigate the protective role of dietary supplementation of Spirulina platensis (SP) against cytotoxic and genotoxic effects of lead nitrate in Clarias gariepinus. Four groups of fishes were used: first group as control which fed on basal diet, second group fed on basal diet and exposed to (1 mg/L of lead nitrate), third group fed on diet containing 0.25% SP and exposed to (1 mg/L of lead nitrate), and fourth group fed on diet containing 0.5%SP and exposed to (1 mg/L of lead nitrate). Fish samples were taken at 2nd and 4th week of exposure. The hematological indices of lead nitrate-exposed group were decreased significantly compared to the control group at 2nd and 4th week of exposure. Lead nitrate caused a significant increase in the percentage of poikilocytosis, micronuclei, and apoptotic cells as well as comet tail length and olive tail moment compared with the control group at 2nd and 4th week of exposure. The highest level of damage was found on 4th week of exposure with all parameters. Dietary inclusion of SP ameliorated these cytotoxic and genetic changes, as well as this amelioration was concentration and time dependent. Consequently, the present study proposed that the addition of SP to the fish diet can be used as a promising protective agent to oppose cytotoxic and genotoxic effects of lead nitrate in aquaculture. Graphical abstract.

Protective effects of a novel pyrazolecarboxamide derivative against lead nitrate induced oxidative stress and DNA damage in Clarias gariepinus.

Pyrazole derivatives display diverse biological and pharmacological activities. The aim of this study is to investigate the antioxidant properties of a novel pyrazolecarboxamide derivative (4-amino-N-[(4-chlorophenyl)]-3-methyl-1-phenyl-1H-thieno [2, 3-c] pyrazole-5-carboxamide) in African catfish, Clarias gariepinus, exposed to 1 mg/L PbNO3. Fish were intramuscularly injected with pyrazole-5-carboxamidederivative according to the following groupings: Group 1 (control), Group 2 (1 mg/L lead nitrate), Group 3 (1 mg/L lead nitrate + 5 mg pyrazole derivative/kg body weight), and Group 4 (1 mg/L lead nitrate + 10 mg pyrazole derivative/kg body weight) for two weeks and four weeks. Lead nitrate (1 mg/L) caused significant elevation of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), creatinine, uric acid, cholesterol, and glucose-6-phosphate dehydrogenase (G6PDH) compared to the control group after two and four weeks of exposure, while serum total lipids, alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) were significantly reduced compared to the control group. Furthermore, levels of antioxidant enzymes superoxide dismutase (SOD) and catalase (CAT) and total antioxidant capacity (TAC) were reduced in group 2 compared to the control group. However, in group 2, hepatic lipid peroxidation (LPO) and DNA fragmentation percentage were significantly increased compared to the control group. Histopathological changes in the liver of lead-exposed groups included marked disturbance of hepatic tissue organization, degeneration of hepatocytes, dilation of blood sinusoids and the central vein as well as necrosis. Injection of pyrazole derivative for two weeks and four weeks reversed alterations in biochemical parameters, antioxidant biomarkers, lipid peroxidation, hepatic DNA damage, and histopathological changes in liver tissue induced by 1 mg/L lead nitrate. This amelioration was higher in response to high-dose pyrazole derivative (10 mg) at the fourth week of exposure, showing concentration-and time-dependency. Overall, the sensitized derivative pyrazolecarboxamide is likely a useful tool to minimize the effects of lead toxicity due to its potent antioxidant activity.

Modulatory effects of green tea extract against the hepatotoxic effects of 4-nonylphenol in catfish (Clarias gariepinus).

The antioxidant role of the green tea (Camellia sinensis) extract (GTE) was examined to remedy the toxic effects of (0.2mgl-1) 4-nonylphenol(4-NP). Biochemical parameters, antioxidant enzymes, liver lipid peroxidation (LPO), DNA fragmentation, and apoptosis as well as histopathology of liver of African catfish Clarias gariepinus were considered. Catfishes were divided into four groups: first group (control), second group (0.2mgl-1 of 4-NP), third group (0.2mgl-1of 4-NP +100mg GTE l-1water), and fourth group (0.2mgl-1 of 4-NP +200mg GTE l-1water). The results showed that significant increments of serum glucose, AST, ALT, total protein, total lipids, cholesterol, G6PDH, and cortisol. Meanwhile, serum acetylcholinesterase, ALP, and LDH were significantly reduced. In addition, antioxidant enzymes (SOD, CAT, GST, and TAC) levels were reduced in 4-NP treated fish compared to control. Also, there were significant increments in hepatic LPO, DNA fragmentation, and apoptotic erythrocytes in 4-NP treated fish compared to control. Liver of 4-NP treated fish showed some histopathological alterations such as, vacuolization in hepatocytes, congestion in central vein, infiltration of mononuclear inflammatory cells, and necrosis as well as depletion of glycogen content of liver. Addition of green tea extract into the water restored the alterations in most of those biomarkers induced by 4-NP. We concluded that, GTE has a protective role against hepatic failure, depletion of antioxidant defense, and genotoxicity induced 4-NP in C. gariepinus.

Hepatoprotective efficacy of Spirulina platensis against lead-induced oxidative stress and genotoxicity in catfish; Clarias gariepinus.

Lead (Pb) is a toxic environmental pollutant that induces a broad range of biochemical and physiological hazards in living organisms. We investigated the possible hepatoprotective effects of Spirulina platensis (SP) in counteracting the Pb-induced oxidative damage. Ninety-six adult African catfish were allocated into four equal groups. The 1st group (control) fed basal diet while the 2nd group (Pb-treated) fed on basal diet and exposed to 1mg Pb(NO3)2/L. The 3rd and 4th groups fed SP-supplemented basal diets at levels of 0.25% and 0.5%, respectively and exposed to Pb. Serum samples were used to analyze hepatic function biomarkers, electrolytes, and oxidant and antioxidant status. Lipid peroxidation and DNA fragmentation were determined in the liver tissues. Pb exposure induced hepatic dysfunction, electrolytes (Na+, K+, Ca+2, and Cl-) imbalance, as well a significant decrease in GSH content, and LDH, AChE, SOD, CAT and GST enzymes activity. SP supplementation reverted these biochemical and genetic alterations close to control levels. This amelioration was higher with 0.5% SP and at the 4th week of exposure, showing concentration- and time-dependency. Thus, the current study suggests that SP could protect the catfish liver against lead-induced injury by scavenging ROS, sustaining the antioxidant status and diminishing DNA oxidative damage. The dietary inclusion of SP can be used as a promising protective agent to counteract oxidative stress-mediated diseases and toxicities.

Endocrine disruption, oxidative stress, and testicular damage induced by 4-nonylphenol in Clarias gariepinus: the protective role of Cydonia oblonga.

Exposure to xenoestrogens like 4-nonylphenol (NP) is recognized by disrupting endocrine functions and causes reproductive dysfunction in male fish. The present study aimed at investigating the 4-nonylphenol propensity to induce oxidative stress and hormonal disturbances in male catfish and at studying the protective role of quince (Cydonia oblonga). To fulfill this aim, catfish Clarias gariepinus were exposed to pure 100 µg/L 4-NP and to quince the leaf extract added to 4-NP, both for 15 days. The 4-NP exposure induced a marked increase in 17ß-estradiol (E2), LH, and cortisol, while thyroid hormone (TSH, T3), testosterone (T), and FSH levels noticeably decreased; however, 4-NP had no effect on T4 level. Moreover, 4-NP exposure was accompanied by histological impairments in testes. Existence of 4-NP was associated with oxidative damage as evidenced by the significant increase (p < 0.05) of the enzymes, superoxidase dismutase (SOD), catalase (CAT), acetylcholinesterase (AchE), glutathione s-transferase, total antioxidant capacity (TAC), and malondialdehyde (MDA). Adding quince was effective to neutralize hormonal levels and to repair the testicular histological alterations. In response to quince remedy, the enzymes AchE and MDA reduced significantly (p < 0.05), while limited or no response was detected for other tested enzymes. Our results concluded that quince can antagonize 4-NP toxicity in catfish, confirming that quince leaf extract displayed antioxidant activities against the toxicity of hazardous chemicals.

Induction of apoptosis and DNA damage by 4-nonylphenol in African catfish (Clarias gariepinus) and the antioxidant role of Cydonia oblonga.

In this study, we assessed the toxic effects of sub lethal concentration (0.1mgl-1) 4-nonylphenol (4-NP) on serum biochemical parameters, liver lipid peroxidation (LPO) and antioxidant enzymes of the African catfish Clarias gariepinus for 14 days and the ability of the quince leaf extract to alleviate the effects of (4-NP). Fish were categorized into four groups: control, exposure to 0.1mgl-1 4-NP, exposure to 0.1mgl-1 4-NP with quince leaf extract (10ml/30L water), and exposure to 0.1mgl-1 4-NP with quince leaf extract (20ml/30L water). 4-NP exposure induced a significant (p<0.05) increase in the levels of glucose, AST, ALT, creatinine, urea, uric acid, cholesterol, and G6PDH as well as, the percentages of hepatic LPO level, DNA fragmentation, and apoptotic erythrocytes (p<0.05). A significant (p<0.05) decrease in alkaline phosphatase (ALP), total protein, albumin, globulin, total lipids, and LDH were also recorded. Liver enzyme activities (SOD, CAT and TAC) were increased. Addition of the quince leaf extract into the water was able to reinstate the alterations in biochemical parameters, antioxidant biomarkers, apoptotic level and hepatic DNA damage induced by 4-NP.

Antioxidant and antiapoptotic activities of Calotropis procera latex on Catfish (Clarias gariepinus) exposed to toxic 4-nonylphenol.

Calotropis procera L. is known as medicinal plant. The Phytochemical analyzes of its latex revealed that it possessed antioxidants, namely terpenes, phenolic compounds and cardenolides, flavonoids and saponins, while tannins, alkaloids and resin were absent in moderate to high concentration. In the present study, the role of latex of Calotropis procera as antioxidant and antiapoptotic was reported. To carry out this aim, fishes were exposed to 100 µg l(-1) 4-nonylphenol as chemical pollutant. The enzymes, superoxidase dismutase, catalase, acetlycholinstrase (AchE), glutathione s-transferase, cortisol, G6PDH) and apoptotic cells increased significantly (p<0.05) accompanied by irregular disturbance of (Na(+), K(+)) ions in the presence of 4-nonylphenol. On the other hand, these enzymes, ions, and apoptotic cells decreased normally and significantly (p<0.05) in the presence of latex. Total phenol content, total capacity antioxidant, reducing power decrease significantly (p<0.05) in the presence of 4-nonylphenol and increase normally in the presence of latex. Latex was used for the first time to protect catfish after 4-nonylphenol exposure. Our study confirms that crude latex of Calotropis procera possessed antioxidant and antiapoptotic activities against the toxicity of 4-Nonylphenol.

Arsenic-induced genotoxicity in Nile tilapia (Orechromis niloticus); the role of Spirulina platensis extract.

Arsenic (As) is one of the most relevant environmental global single substance toxicants that have long been regarded as a carcinogenic and genotoxic potential. In this respect, we evaluated the cytogenetic effect of arsenic exposure in Nile tilapia (Oreochromis niloticus), in terms of erythrocyte alteration, apoptosis, and induction of micronuclei. Spirulina platensis (SP) is a filamentous cyanobacterium microalgae with potent dietary phytoantioxidant, anti-inflammatory, and anti-cancerous properties supplementation. The protective role of Spirulina as supplementary feeds was studied in Nile tilapia (O. niloticus) against arsenic-induced cytogenotoxicity. Four groups were assigned as control group (no SP or As), As group (exposed to water-born As in the form of NaAsO2 at 7 ppm), SP1 (SP at 7.5% + As at the same level of exposure), and SP2 (SP at 10% + As at the same level of exposure). As-treated group had a significant increase in all cytogenetic analyses including erythrocyte alteration, apoptosis, and induction of micronuclei after 2 weeks with continuous increase in response after 3 weeks. The combined treatment of Spirulina at two different concentrations of 7.5 and 10% had significantly declined the induction of erythrocyte alteration, apoptosis, and micronuclei formation induced by arsenic intoxication.

Use of hematological parameters to assess the efficiency of quince (Cydonia oblonga Miller) leaf extract in alleviation of the effect of ultraviolet--A radiation on African catfish Clarias gariepinus (Burchell, 1822).

The present study aimed to elucidate the negative impacts of UVA on some biochemical and hematological variables of the economically important African catfish, Clarias gariepinus and investigates the putative role of quince (Cydonia oblonga Miller) leaf extract in protection and/or alleviation of such negative impacts. Changes in the hematological and blood biochemical values often reflect alteration of physiological state. Blood parameters can be useful for the measurement of physiological disturbances in stressed fish and thus provide information about the level of damage in the fish. We found a significant (P<0.05) decrease in the red blood cell counts, hemoglobin and hematocrit in the groups exposed to UVA compared to the control groups. Exposure to UVA induced marked red cell shrinkage (increased mean cell hemoglobin concentration) and showed an elevation in mean cell volume and mean cell hemoglobin in the blood of the exposed fish compared to the control. A significant (P<0.05) reduction in the total white blood cells was recorded in the exposed fish compared to the control. The biochemical parameters (blood glucose, total plasma protein, blood cholesterol, plasma creatinine, aspartic amino transferase and alanine amino transferase) exhibited a significant increase in the blood of fish exposed to UVA. Methanolic extract of quince leaf before ripening of the fruits was analyzed by GC/MS. To investigate the biological impact of this extract and its biologically active components, this extract was tested for its putative role in alleviation of UVA effect on catfish. Quince leaf extract had the ability to prevent hematotoxic stress induced by UVA and resulted in enhancement of the immune system of catfish represented by significant (P<0.05) increase in the number of white blood cells and lymphocytes of the catfish. Quince extract also protected the red blood cells from UVA damage. To our knowledge this is the first report of the effect of quince leaf extract on an aquatic organism.