Alaa El-Din H. Sayed

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.

Effects of 4-nonylphenol on blood cells of the African catfish Clarias gariepinus (Burchell, 1822).

In the present work, the destructive effects of the 4-nonylphenol on one of the most economically important Nile fishes, namely African catfish (Clarias gariepinus) were studied. Apoptosis, erythrocytes alterations, micronucleus test and blood parameters count were used as biological indicators to detect those effects. After exposure to sublethal concentrations of 4-nonylphenol (0, 0.05, 0.08 and 0.1mg/l), apoptotic red blood cells with many malformations and micronucleated erythrocytes were recorded. Decrease in the blood parameters such as red blood cells (RBCs), hemoglobin (Hb), package cell volume (PCV), mean corpuscular hemoglobin concentration (MCHC), platelets, white blood cells (WBCs), lymphocytes, basophils, monocytes and increase in mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), neutrophils, eosinophils indicated the negative effects of 4-nonylphenol. It was concluded that, the 4-nonylphenol caused genotoxicity in erythrocytes with many malformations in shape and number indicated with other blood parameters.

Reproductive biomarkers to identify endocrine disruption in Clarias gariepinus exposed to 4-nonylphenol

The present study investigated the hormones concentrations and gonads alterations of Clarias gariepinus caused by sublethel concentrations of 4-nonylphenol (0, 0.05, 0.08 and 0.1 mg/l). The changes in the activities of the hormones after exposure to these sublethel doses of 4-nonylpenol referred to endocrine disruption in Clarias gariepinus in association with histopathological changes in reproductive tissues. The levels of thyroid stimulating hormone (TSH), triiodothyronine (T3), total thyroxine (T4), follicle stimulating hormone (FSH), luteinizing hormone (LH) and testosterone concentrations significantly decreased (P<0.05) in the treated fish in comparison with control. 17-β-estradiol increased significantly (P<0.05) with 4-nonylphenol concentrations increase. Reduction in the gonadosomatic index was evident with increase of sublethal doses of 4-nonylphenol. The histopathological changes of NP-treated were recorded in gonads of Clarias gariepinus reflecting their sensitivity to NP-estrogenic like effects.

Ultraviolet radiation-A (366 nm) induced morphological and histological malformations during embryogenesis of Clarias gariepinus (Burchell, 1822)

Exposure to ultraviolet radiation has been associated with variety effects in many organisms ranging from molecular and tissue damage to population level effects. The exposure of embryos of the catfish, Clarias gariepinus (Burchell, 1822) to 366nm UVA at different doses 15, 30 and 60 min resulted in the hatching time delayed to 29 h-post-fertilization stage (29 h-PFS) in comparison with normal hatching time of 22h-PFS at 29 degrees C. In embryos exposed to 15 min/UVA, 30 min/UVA and 60 min/UVA the total percentage of hatched embryos/fertilized eggs were 90%, 89% and 85%, respectively, while in control was 95% at 29 h-PFS. The total percentage of mortality/ hatched embryos were (1-14)%, (2-22)%, (2-23)% and (3-40)% for control, 15 min, 30 min and 60 min groups, respectively, at 40 h-PFS. Also as a result some morphological malformations; (yolk sac oedema, body curvature, fin blistering, and dwarfism) were revealed. These destructive effects were also confirmed by histopathological changes in gills, eyes, intestinal tract, spinal cord, notochord, liver, skin and kidney. The results confirm that exposure to UVA caused an exposure time-dependent delay in hatching rate and reduced the percentage of the hatched embryos but the mortality rate increased with increase of the exposure time to UVA.

The protective role of quince leaf extract against the adverse impacts of ultraviolet--a radiation on some tissues of Clarias gariepinus (Burchell, 1822).

In the present study the protective role of quince leaf extract against the adverse impacts of ultraviolet radiation-A (UVA) on some tissues of Clarias gariepinus (Burchell, 1822) was considered. Fishes were classified into four groups: control, UVR-treated group (for 3days/for 3h/day), UVR-treated group (for 3days/for 3h/day) with adding 10ml of quince extract, and UVR-treated group (for 3days/for 3h/day) with adding 20ml of quince leaf extract. Blood smears and sections of the liver, and skin were processed routinely for H & E paraffin embedding technique. Some UVA-induced malformations were recorded in the red blood cells including crenated cells (Cr), Acanthocytes (Ac), tear drop-like cells (Tr) and sickle cells (Sk). Also, UVA-induced disorganization of the normal architecture of hepatic tissues with lipidosis was evident. Hypertrophy and vacuolated club cells were recorded in skin exposed to UVA. In conclusion, quince leaf extract has a valuable antioxidant protective role to prevent and/or repair the histopathological changes induced by UVA.

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.

Histopathological Alterations in some Body Organs of Adult Clarias gariepinus (Burchell, 1822) Exposed to 4-Nonylphenol

Endocrine-disrupting chemicals (EDCs) include synthetic and naturally occurring chemicals that affect the balance of normal functions in animals (Razia et al. 2006). It has been found that exposure to natural and synthetic estrogenic chemicals may adversely affect wildlife and human health (Colborn et al. 1993). In vitro exposures (Soto et al. 1992; Soto et al. 1994; Toomey et al. 1999) have confirmed the effects of EDCs on tissue structure and cellular processes. Nonylphenol ethoxylates (NPEs) are EDCs which are used globally in the production of plastics, pesticides, and cleaning products and are present in sewage effluents around the world (Talmage, 1994). It has been reported that NP is the most important degradation product of NPEs because of its enhanced resistance towards biodegradation, toxicity, ability to bioaccumulate in aquatic organisms, and estrogenicity (Ahel et al. 1994). NP is found in surface waters, aquatic sediments, and ground water (Bennie, 1999; Talmage, 1994) and it is estrogenic in various aquatic animals (Nimrod and Benson, 1996; Talmage, 1994; Servos, 1999).

Apoptosis and morphological alterations after UVA irradiation in red blood cells of p53 deficient Japanese medaka (Oryzias latipes).

Morphological alterations in red blood cells were described as hematological bioindicators of UVA exposure to investigate the sensitivity to UVA in wild type Japanese medaka (Oryzias latipes) and a p53 deficient mutant. The fewer abnormal red blood cells were observed in the p53 mutant fish under the control conditions. After exposure to different doses of UVA radiation (15min, 30min and 60min/day for 3days), cellular and nuclear alterations in red blood cells were analyzed in the UVA exposed fish compared with non-exposed controls and those alterations included acanthocytes, cell membrane lysis, swollen cells, teardrop-like cell, hemolyzed cells and sickle cells. Those alterations were increased after the UVA exposure both in wild type and the p53 deficient fish. Moreover, apoptosis analyzed by acridine orange assay showed increased number of apoptosis in red blood cells at the higher UVA exposure dose. No micronuclei but nuclear abnormalities as eccentric nucleus, nuclear budding, deformed nucleus, and bilobed nucleus were observed in each group. These results suggested that UVA exposure induced both p53 dependent and independent apoptosis and morphological alterations in red blood cells but less sensitive to UVA than Wild type in medaka fish.

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.

Nuclear and cytoplasmic changes in erythrocytes of p53-deficient medaka fish (Oryzias latipes) after exposure to gamma-radiation

Previous studies have examined the effects of gamma-radiation on Japanese fish, in particular medaka (Oryzias latipes). In the present work, alterations in erythrocytes were recorded as haematological bio-indicators of exposure to gamma-radiation. After exposure of medaka fish to two different doses of radiation (2 Gy and 10 Gy), many malformations in red blood cells were observed in the irradiated fish compared with control fish. These malformations included acanthocytes, crenated cells, amoeboid cells, and sickle cells. More malformations were seen at the higher radiation dose. No micronuclei were seen in any group, but nuclear abnormalities were observed. We conclude that gamma-radiation causes morphological malformations of erythrocytes and is harmful to medaka fish.