الفهرس 
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Abstract
In the present work specimens of catfish, Clarias
gariepinus were used to investigate the adverse effects of the
sublethel concentrations of 4-nonylphenol (0.05, 0.08, and 0.1
mg/l) on hematological parameters, biochemical variables,
hormonal disruption and genotoxicity. The histopathological
changes of gills, skin, liver and kidney were also studied.
Many advanced techniques as Micronucleus test, apoptosis
detection, SDS-Page, RAPD-PCR, TEM, and SEM were used to
determine the effects of 4-nonylphenol on Clarias gariepinus.
Also, incubation period, fertilization rate, hatching rate and
mortality rate were used to investigate the effects of 4-nonylphenol
on embryos of Clarias gariepinus.
The results can be summarized in the following items
1. Heamatological parameters
• Apoptotic, micronucleated and altered erythrocytes
The number of apoptotic, micronucleated and altered
erythrocytes were increased significantly (p< 0.05) with
increase of 4-nonylphenol concentrations.
• Heamatological parameters changes
• The number of R.B.Cs, Hemoglobin content,
hematocrite, MCHC, Blood Plateletes, white blood
cells, lymphocytes, monocytes and basophils were
significantly decreased at p<0.05 after exposure to 4-
nonylphenol • MCV, MCH, neurtophils and eosinophils were
increased significantly (P<0.05) after exposure to 4-
nonylphenol.
• Also, reduced swimming with a very weak appetite
and haemorrahgic patches with anoxia were
observed.
2. Biochemical blood parameters
• ALT, AST, G6PDH, total protein, uric acid, glucose,
cholesterol, creatinine, K+, and Fe +2 were increased
while LDH, Hco3
-, Na+, Cl-, Cu+2 and Ca+2 were
decreased under the effects of 4-nonylphenol.
• Variability numbers and molecular weights of protein
fractions under the effects of 4-nonylphenol were
observed.
3. Hormonal activities disruption
• TSH, T3, T4, FSH and testosterone were significantly
decreased at p <0.05 while LH, and 17-β estradiol
were significantly increased after exposure to 4-
nonylphenol
• The sex has a significant effect on T3, LH,
testeosterone, 17-β estradiol but no effect of sex on
TSH, T4 and FSH.
4. Reproductive system alterations
a. Gonado-somatic index • The GSI decreased significantly p< 0.05 in
fishes with 4- nonylphenol in comparison with
control fishes
• The decrease in GSI was significant with
increase of 4- nonylphenol concentrations.
b. Testis alterations
Histopathological changes were recorded after
exposure to different concentrations of 4-nonylphenol
as
• Abnormal distribution of spermatozoa
• Degeneration of spermatozoa
• Rupture of interlobular tissue
• Vacuolation around seminiferous tubules
• Few number of Sertoli cell with nuclei
hypertrophy
• Dead sperms and decrease of spermatogonia
nests.
c. Ovaries alterations
Treatment fishes with (0.05, 0.08 and 0.1mg/l) of
4-nonylphenol for 15 days showed histopathological
changes in the ovary as
• Retraction of the cytoplasm from the follicular
cells
• Irregular shape of oocytes and discontinuous
follicular layer
• Yolk granules and zona radiate were broken • Oocyte size reduction with irregularities in
shape
d. Ultra-structure of the spermatozoa
The fish treated with 0.1 mg/l of 4-nonylphenol
for 15 days exhibited malformed sperms with many
malformations such as
• Vacuolation around axoneme
• Reduction in the nucleus size
• Damaged mitochondria and abnormal flagellum
5. Genotoxicity
a. Total DNA concentration
* Significant decrease in the concentrations of
genomic DNA with increase of 4-nonylphenol
doses in adults and embryos of catfish Clarias
gareiepinus
b. Selection of PCR primer
* Ten different random primers were used and
only one gave 4-9 bands and evidenced
significant difference between control and
treated fishes
c. Analysis of 4-nonylphenol-induced genotoxicity
* The profile of RAPD-PCR of fish exposed
to 4-nonylphenol showed disappearance and
appearancne of some bands with difference in
molecular weight
* Dendrogram showed the relationship
among applied doses of 4-nonylphenol on adults fishes and among the 4-nonylphenol
and the developmental stages
6. Histopathological changes
• The number of dead fish was 1.2, 3.75, 5.5 and
8 % for control, 0.05, 0.08, and 0.1 mg/l of 4-
nonylphenol respectively.
• Lesions were observed in the gills, skin, kidney
and liver of sampled fish for all 4-nonylphenol
at all exposure concentrations and durations as
following:-
a. Gills
• Epithelial lifting and hyperplasia
• Edema of epithelium
• Deformed secondary lamellae with diffusion
• Desquamation and necrosis
• Increase in number and size of chloride cells
• Increase in number of mucous cell
b. Skin
• Enlarged alarm cells
• Rupture of epithelial cells
• Necrosis, granuled and fat cells in epithelium
c. Kidney
• Edema in renal tubules
• Degeneration of renal tubules
• Rupture of bowman’s capsule
• Hypertrophy of the glomerulus • Shrinkage of the glomerulus
• Presence of necrosis and pyknosis
• Dilated blood vessels
• Increase of melanomacrophages
• Obliterated Bowman’s space
d. Liver
• Degeneration of hepatocytes
• Lymphocytic aggregation
• Increase of melanomacrophages
• Rupture of hepatocytes membranes
• Presence of pyknotic cells
• Presence of necrotic areas
• Hyperplasia of Küffer cells
• Pigment diffusion
e. Electron microscope examination of hepatocyte
Hepatocytes of fish exposed to 4-nonylphenol
shows many alterations as:
• Swollen or hypertrophied hepatocytes with dense
bodies
• Karyolsis in nuclei
• Damaged mitochondria
• Rarified cytoplasm
• Shrunken and indented nuclei
• Blood sinusoids collapsation
• Apoptotic and vacuolated hepatocytes
• Increase in number of lysosomes and fat DROPs 7. Morphological malformations
a. Fertilization rate, incubation period, hatching rate, and
mortality rate were affected by exposure to 4-
nonylphenol as:
• Fertilization rate increased
• Incubation period increased
• Hatching rate decreased
• Mortality rate increased
b.Many gross morphological malformations were
recorded in groups exposed to sublethel concentrations of
4-nonylphenol as
• Yolk sac oedema
• Body curvature (lateral curvature, dorsoventral
curvature and ventrodorsal curvature)
• Fin blistering
• Irregular head shape
• Dwarfism
• Pericardial oedema
• Collapsed tail
• Egg shrinkage
c. Histopathological changes were recorded in the spinal
cord and notochord in the post- hatching stages after
exposure to 4-nonylphenol.