Reproductive Toxicity and Recovery Associated With 4-Non-ylphenol Exposure in Juvenile African Catfish (Clarias garepinus).

Although, the effects of 4-non-ylphenol (4-NP) on fish's reproductive hormones were assessed in several studies using adult models, however, the effect of this endocrine disruptor on immature fish's reproductive hormones was not addressed commonly. We aimed to study the apoptosis induction, hematotoxicity, reproductive toxicity, and the recovery associated with 4-NP exposure in juvenile African catfish [Clarias garepinus) using some hormones [17ß-estradiol (E2), testosterone (T), follicle-stimulating hormone (FSH), and luteinizing hormone (LH)] and gonad histology as biomarkers. The toxic effects of 4-NP have been studied in many animal models, but there is still limited knowledge about the dose-dependent damage caused by 4-NP exposure in juvenile Clarias gariepinus. A healthy juvenile C. gariepinus was categorized into four groups (n = 3/group; three replicates in each group). The first group was the control, and the other three groups were subjected to 4-NP concentrations as 0.1, 0.2, and 0.3 mg/L for 15 days; they were left for a recovery period of another 15 days. The reproductive hormones of C. gariepinus exposed to 4-NP for 15 days exhibited significant variations between the treatment groups and the control (P < 0.05), which were evident in E2 and T-values, whereas FSH, LH, total protein, and lipid peroxidation values showed non-significant differences among all groups at P < 0.05. Such a situation referred to the fact that the 15-day recovery period was insufficient to remove the impacts of 4-NP doses in concern. The trend of dose-dependent increase/decrease was recorded for T, E2, FSH, and LH. The histopathological alterations of 4-NP treated in gonad tissues were recorded in juvenile C. gariepinus, reflecting their sensitivity to 4-NP estrogenic-like effects. Overall, our results investigate that recovery has improved the reproductive toxicity caused by 4-NP in juvenile C. garepinus. Significant variations between the treated groups and the control group (P < 0.05) were evident in hematological parameters except for hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and mean corpuscular hemoglobin concentration (MCHC). The parameters exhibiting significance decreased with such increased doses [red blood cells (RBCs), hematocrit (Hct), and white blood cells (WBCs)]. Similar patterns of significant variations toward the increase or decrease were recorded following the 15-day recovery period. Apoptotic frequency in erythrocytes and brain cells has increased significantly with increased 4-NP exposure, indicating that 4-NP caused cytotoxic effects, such as apoptosis in a dose-dependent manner. However, these cellular alterations greatly decreased after the 15-day recovery period.

Comparative study of two carnivorous fish (Parupeneus forsskali and Thalassoma klunzingeri) from the Red Sea: Hemato-biochemical parameters and cellular characterization.

Although, the Red sea is highly rich with fish fauna but still the information known is so limited for the researchers especially about the fish physiology baselines. So, in the present study we investigated the heamto-biochemical parameters and cell characterization of two fish having the same feeding habitats. Fish specimens of Red Sea goatfish (Parupeneus forsskali) and Klunzinger's wrasse (Thalassoma klunzingeri) were captured from Hurghada, Egypt. Haematological and biochemical analysis as well as blood cells characterization were performed. The morphological and cytochemical aspects of peripheral blood cells of the two species were studied by light microscopy. Thalassoma klunzingeri showed lower Hct and Hb values and RBCs count. Fusiform to spindle shape thrombocytes were found only in the blood of Parupeneus forsskali while spiked thrombocytes were found only in the blood of Thalassoma klunzingeri. This investigation may be helpful as a tool to monitor the health status of the two species and will be used as biomarkers for clinical pathology.

Hepatotoxic effects of silver nanoparticles on Clarias gariepinus; Biochemical, histopathological, and histochemical studies.

The current study investigates the hepatotoxic effects of two acute doses of silver nanoparticles (AgNPs) and silver nitrate (AgNO3) on African catfish (Clarias garepinus) using biochemical, histopathological, and histochemical changes and the determination of silver in liver tissue as biomarkers. AgNPs-induced impacts were recorded in some of these characteristics based on their size (20 and 40 nm) and their concentration (10 and 100 µg/L). Concentrations of liver enzymes (Aspartic aminotransferase; AST, Alanine aminotransferase; ALT), alkaline phosphatase (ALP), total lipids (Tl), Glucose (Glu) and Ag-concentration in liver tissue exhibited a significant increase under stress in all exposed groups compared to the control group. The total proteins (Tp), albumin (Al), and globulin (Gl) concentrations exhibited significantly decrease in all treated groups compared to the control group. At tissue and cell levels, histopathological changes were observed. These changes include proliferation of hepatocytes, infiltrations of inflammatory cells, pyknotic nuclei, cytoplasmic vaculation, melanomacrophages aggregation, dilation in the blood vessel, hepatic necrosis, rupture of the wall of the central vein, and apoptotic cells in the liver of AgNPs-exposed fish. As well as the depletion of glycogen content in the liver (feeble magenta coloration) was observed. The size and number of melanomacrophage centers (MMCs) in liver tissue showed highly significant difference in all exposed groups compared to the control group. Recovery period for 15 days led to improved most alterations in the biochemical, histopathological, and histochemical parameters induced by AgNPs and AgNO3. In conclusion, one can assume liver sensitivity of C. garepinus for AgNPs and the recovery period is a must.

Silver nanoparticle-induced nephrotoxicity in Clarias gariepinus: physio-histological biomarkers.

The present study investigates the nephrotoxic effects of two acute doses of silver nanoparticles (AgNPs) and silver nitrate (AgNO3) on the African catfish, Clarias gariepinus, using biochemical, histochemical, and histopathological changes as biomarkers. AgNP-induced impacts were recorded in some of these characteristics on the bases of their size (20 and 40 nm) and concentration (10 and 100 µg/L) but no significant interaction between size and concentration. AgNO3 had low significant adverse effects on some parameters in comparison with those impacts of AgNPs. The concentrations of creatinine and uric acid exhibited different significant variations under stress in all exposed groups compared with those in the control group. On the tissue and cell levels, histopathological changes were observed. These changes include hypertrophies of glomeruli, proliferation in the haemopoietic tissue, dissociation in renal tubules, shrinkage of glomerulus, hydropic degeneration, dilatation of renal tubules, aggregation of melanomacrophages, rupture of Bowman's capsule, and the glomerular tuft and dilatation of Bowman's space. In more severe cases, the degenerative process leads to tissue necrosis in the kidney of AgNP-exposed fish as well as carbohydrate depletion; a faint coloration was also observed in the brush borders and basement membrane with a large amount of connective tissue fibers around the blood vessels and the renal tubules. Recovery period for 15 days led to improvement of most of the alterations in biochemical, histopathological, and histochemical parameters induced by AgNPs and AgNO3. In conclusion, one can postulate on the sensitivity of the kidney of C. gariepinus to AgNPs and recovery strategy is a must.

Cytotoxic and hemotoxic effects of silver nanoparticles on the African Catfish, Clarias gariepinus (Burchell, 1822).

The present study investigates the hemotoxic and cytotoxic impacts of two acute doses of silver nanoparticles (AgNPs) on the African catfish, Clarias garepinus in comparison to the impact of AgNO3 and the control fish. AgNPs-induced impacts were recorded on some biological and hematological indices of that species on the bases of their size (20 and 40 nm) and concentration (10 and 100 µg) but no significant interaction. AgNO3 had very low impact on these indices in comparison to AgNPs. Recovery period for 15 days was found to be valid to remove AgNPs and AgNO3 toxicity for most indices. The condition factor exhibited stability under stress whereas the hepatosomatic index was more sensitive to AgNPs. The AgNPs-induced hematological changes recorded were corresponding with different blood cell alterations which increased in frequency from the control and AgNO3 to 40 nm/100 µg; such blood cell alterations disappeared to great extent after recovery period of 15-days in a reverse order.

The microstructure of buccal cavity and alimentary canal of Siganus rivulatus: Scanning electron microscope study.

The microstructure of the oral cavity and alimentary canal of herbivorous fish Siganus rivulatus collected from the Red Sea were investigated by using scanning electron microscope (SEM). The results showed that S. rivulatus has three types of teeth, tri-cusped, bi-cusped, and papilliform. A taste bud (Type I) was recorded in the oropharyngeal cavity. Characteristic styles of microridges on the cell's surface inside the buccal cavity were recorded. Also, the distribution of the mucous cells in the lining of the mouth cavity, alimentary canal was observed. Mucosal folds along the distinct parts of alimentary canal, showed characteristic pattern which was complex in the intestinal mucosa. The results concluded that there are characteristic microstructures according to feeding habitat compared with other bony fishes.

SDS-PAGE technique as biomarker for fish toxicological studies.

Although many studies on the hematological and biochemical parameters in fishes have been done, still there are some shortage in the estimation and evaluation of the baseline's values of marine and freshwater fishes. Recently, the use of hematology and biochemistry of fishes in toxicology, aquaculture, environmental pollution, feeding, and antioxidants studies has been increased. In this study we introduced the importance of those parameters and their importance as biomarkers in fish toxicology from previous literature and as new findings. Hemato-biochemical parameters were widely used in fish toxicological studies. Many researches have used the protein electrophoresis as a valid tool to determining intra and inter-specific variation among species. Protein profile was extensively used in determining the health of fish, as indicators of anemia or other diseases provide information about the existence of the disease, and in the diagnosis of disease. So, to carry out the aim of this study, we reported one of the more advanced techniques used SDS-PAGE as molecular biomarker for protein profile analysis in fish with shedding the light on the importance of hematological and biochemical parameters in fish toxicological studies.

Surface architecture of the oropharyngeal cavity and the digestive tract of Mulloidichthys flavolineatus from the red sea, Egypt: A scanning electron microscope study.

Surface architecture of the buccal cavity and the surface organization of the luminal mucosa of the oesophagus, stomach, and intestine of the carnivorous fish M. flavolineatus from the Red Sea were studied by using SEM. The results revealed that M. flavolineatus has four kinds of teeth; curved-blunt, wedge-shaped, flattened crowns, molariform and papilliform. Three types of taste buds (type I, II and III) were recorded in the oropharyngeal cavity. It was observed that taste buds and teeth are co-located in the pharyngeal region. Characteristic patterns of microridges of the surface cells in the oral cavity and oesophagus were observed. Mucous cells are distributed in the lining of the mouth cavity, oesophagus, stomach, and intestine. Characteristic patterns of mucosal folds throughout the alimentary canal, concerning oesophagus, stomach, and intestine were revealed. Numerous gastric pits, which represents the emergence of the gastric glands, were recorded in the anterior and middle regions of the stomach. Complex patterns of the folds and mucous cells were recorded in the intestinal mucosa. These results were discussed with other teleost fishes.

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-ß-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.

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.

Effects of cadmium on some haematological and biochemical characteristics of Oreochromis niloticus (Linnaeus, 1758) dietary supplemented with tomato paste and vitamin E.

The present study investigates the potential protective effects of tomato paste (9 mg/kg-lycopene) in comparison with vitamin E (50 mg/kg) against the impacts of cadmium (Cd) toxicity (4.64 mg/l: » of 96 h LC50) on fishes Cd exposed for 15 and 30 days. Cd impacts were evaluated in terms of biological, haematological and biochemical characteristics. Cd significantly induced free radicals in serum and liver. The activities of aspartate aminotransferase and alanine aminotransferase in serum were significantly increased due to Cd. Treatment with Cd caused a significant increase in Lipid peroxidation and DNA fragmentation in liver tissue and serum glucose and total lipid. On the other hand, Cd significantly led to decline in serum total protein, blood haemoglobin, red blood cell count, haematocrit value, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration. Dietary supplementation with vitamin E and/or tomato paste to Cd-exposed fish declined significantly the increased lipid peroxidation and DNA fragmentation in liver tissue and the increased aspartate aminotransferase, alanine aminotransferase, glucose and total lipid in serum to the normal condition. This supplementation also significantly increased the declined serum total protein, blood haemoglobin, red blood cell count, haematocrit value, mean corpuscular volume, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration to the normal state. Cd impacts and tomato paste/or vitamin E supplementations did not reflected on the condition factor of the fish. These findings demonstrated the beneficial diet supplementation of tomato paste phytonutrients and vitamin E in counteracting the harmful effects of Cd on the characters investigated.

Effects of ultraviolet A on the activity of two metabolic enzymes, DNA damage and lipid peroxidation during early developmental stages of the African catfish, Clarias gariepinus (Burchell, 1822).

Many ultraviolet-A (UVA)-induced biochemical and physiological changes are valid as biomarkers using aquatic species for detection of the degree of stress. Changes in the concentration and activities of enzymes, such as glucose-6-phosphate dehyderogenase (G6PDH), lactate dehyderogenase (LDH), DNA damage and lipid peroxidation (LPO), can be used as biomarkers to identify possible environmental contamination in fish. This study aimed to investigate the impact of UVA on the activity of the selected enzymes, DNA damage and LPO during early developmental stages of the African catfish Clarias gariepinus. Embryo hemogenates were used for measurements of G6PDH, LDH, DNA damage and LPO concentrations and activities spectrophotometrically at 37 degrees C. The normal ontogenetic variations in enzyme activities, DNA damage and LPO of the early developmental stages (24-168 h-PFS; hours-post fertilization stage) were studied. There was a significant decrease in the activity of G6PDH till 120 h-PFS. Then after 120 h-PFS, the activity of such enzymes insignificantly increased toward higher stages. The LDH activity was recorded with a pattern of decrease till 96 h-PFS, followed by a significant increase toward 168 h-PFS. The polynomial pattern of variations in DNA damage and LPO was also evident. The patterns of the enzyme activities, corresponding DNA damage and LPO of the early ontogenetic stages under the influence of three different UVA doses (15, 30 and 60 min), were recorded. The pattern of variations in G6PDH activity in UVA-induced groups was similar to that of the control group with variation in the magnitude of such activity. In all treated groups, LDH activity decreased till 96 h-PFS, then increased till 168 h-PFS. Within each of the embryonic stages, the increase in UVA led to a significant increase in DNA damage. A significant increase in lipid peroxidation under UVA doses was recorded. The variability in number and molecular weight of proteins under exposure to UVA was evident, reflecting some of the genetic and transcriptional changes during exposure and development.

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.

Acute effects of ultraviolet-A radiation on African catfish Clarias gariepinus (Burchell, 1822).

In the present work, the destructive effects of Ultraviolet-A radiation on the African Catfish, Clarias gariepinus was revealed in terms of total protein, cholesterol, glucose, hemoglobin and erythrocytic indices, differential blood cell counting, heamatocrite, creatinine level, Aspartic Amino Transferase, Alanine Amino Transferase and Alkaline Phosphatase. These destructive effects were also confirmed by histopathological changes in liver, blood corpuscles and skin.