Estudios ultraestructurales de las variaciones morfológicas de la superficie y envolturas del huevo de pez-gato africano Clarias gariepinus (Burchell, 1822) antes y después de la fertilización, con una discusión de los mecanismos de fertilización

Much of the existing knowledge of the mechanisms involved in teleost fertilisation is based on a few small model species that have no commercial value. Research is therefore urgently required to address mechanisms involved in fertilisation in species of great commercial value. In this study, the ultrastructural morphological variations in the surface of the egg of Clarias gariepinus were recorded before and after fertilisation by using electron microscopy. The outer surface of the unfertilised egg was smooth, whereas the fertilised egg acquired a network of projections on the vegetal hemisphere. Moreover, different patterns of ornamentation on the egg surface were evident. This pattern of ornamentation varied with the progress of embryonic development. The micropyle of the C. gariepinus egg consisted of a funnel-shaped vestibule, from the bottom of which a cylindrical micropylar canal extended. The micropylar canal decreased in diameter after completion of fertilisation, forming a micropylar disc. The sperm behaviour on the egg surface was oriented towards any depression on the chorion surface. The chorion of ovulated eggs consisted of one layer. After fertilisation the chorion was differentiated into three layers: the double-layered coat, the zona radiata externa and the zona radiata interna. Four protein subunits of the chorion of C. gariepinus were identified by SDS-PAGE. IR-spectra obtained from C. gariepinus chorion revealed that the vibration of chorion proteins exhibited different weak activities in the IR-spectra with minor difference between pre- and post-fertilisation chorion proteins.Gran parte del conocimiento existente sobre los mecanismos de fertilización en teleósteos se basa en unas pocas especies modelo, que no tienen valor comercial. Por ello, se requiere investigación dirigida a conocer los mecanismos involucrados en la fertilización de especies de gran valor comercial. En el presente estudio, mediante el uso del Microscopio Electrónico, se obtuvieron imagines de las variaciones en la morfología ultraestructural de la superficie de los huevos de Clarias gariepinus antes y después de la fertilización. La superficie externa de los huevos infertilizados era lisa, mientras que la de los huevos fertilizados adquiría una red de proyecciones en el hemisferio vegetativo. Además, fueron evidentes diferentes patrones de ornamentación de la superficie del huevo. Este patrón de ornamentación varió a lo largo del desarrollo embrionario. El micropilo de los huevos de C. gariepinus consistió en un vestíbulo con forma de embudo desde la base del cual se extendía un canal micropilar cilíndrico. El diámetro de dicho canal disminuía tras la finalización de la fertilización, formándose el disco micropilar. El comportamiento del esperma en la superficie de los huevos se orientaba hacia cualquier depresión de la superficie del corion. El corion de los ovocitos consistió en una sola capa. Tras la fertilización, el corion se diferenció en tres capas: la capa double-acodada, la zona radiada externa and y la zona radiada interna. Mediante SDS-PAGE se identificaron cuatro subunidades de proteínas en el corion de los huevos de C. gariepinus. Los espectros IR obtenidos del corion revelaron que la vibración de las proteínas del corion exhibía diversas débiles actividades en el espectro IR con menores diferencias entre pre- y post-fertilización proteínas del corion

Erythrocytes alterations of monosex tilapia (Oreochromis niloticus, Linnaeus, 1758) produced using methyltestosterone

AbstractThe present study aims to investigate the effects of methyltestosterone on monosex farmed tilapia, Oreochromis niloticus by detection of apoptosis, micronucleus and alterations of erythrocytes. Fishes were obtained from four localities (Assiut as a control and Beheira, Alexandria and Kafr EL-Sheikh; three farms from each governorate as farmed monosex produced using methyltestosterone). Blood smears were processed for Hematoxylin and eosin technique. The major alterations recorded in the red blood cells were as swelled cells (Sc), tear drop-like cells (Tr), and sickle cells (Sk). Also, a significant difference (P⩽0.001) between three governorates and Assiut was recorded in the micronucleus test, apoptosis and altered erythrocytes. These alterations are considered as an indication for performance and health of fish in the monosex culture medium indicating the side effects of overdose induction of MT

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°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

Effects of lead nitrate on the activity of metabolic enzymes during early developmental stages of the African catfish, Clarias gariepinus (Burchell, 1822)

Glucose-6-phosphate dehydrogenase (G6PDH), lactate dehydrogenase (LDH) and pyruvate kinase (PK) are key metabolic enzymes. G6PDH has been used as a biomarker of pollution-induced carcinogenesis in fish. LDH has been used as marker of lesions in toxicology and clinical chemistry, and PK catalyses the conversion of phosphoenol pyruvate to pyruvate, with regeneration of ATP. The effect of different concentrations of lead nitrate on the activity of these enzymes in two different early ontogenetic stages (embryonic and free embryonic stage) of the African catfish Clarias gariepinus was investigated. Embryo homogenates were used for measurements of G6PDH, LDH and PK activity spectrophotometrically at 340 nm and 25°C. The ontogenetic variations of the three enzymes during early ontogeny, from the 30 h to the 168 h post-fertilisation stage (PFS) (beginning of exogenous feeding), were studied. There was a significant decrease in activities of all three enzymes from 30 h-PFS to 96 h-PFS, followed by a significant increase in G6PDH and LDH. PK showed insignificant fluctuations in activity. Different patterns of enzyme activities were recorded due to exposure to different lead nitrate concentrations (100 ¿g/l, 300 ¿g/l and 500 ¿g/l). In the pre-hatching stage (30 h-PFS) the activity of the three enzymes increased at exposure to 100 ¿g/l lead nitrate and then decreased with increasing dose. In the post-hatching stages (48 h-PFS¿168 h-PFS) G6PDH activity increased and LDH activity decreased with increasing lead concentrations. Unlike G6PDH and LDH, the PK enzyme fluctuated during the post-hatching stages and did not reveal a specific trend of response (increase or decrease) with increasing lead concentrations. Therefore, the measurement of G6PDH and LDH activities, but not PK activity, could be useful biomarkers of intoxication to reveal the embryotoxic potential of lead nitrate in fish embryos. The post-hatching stages of the African catfish are more sensitive than the pre-hatching stage (30 h-PFS) is, probably due to the protective capacity provided by the hardened chorion. The interaction and the main effects of age and lead doses were found to be highly significant, referring to the great impact of lead on these enzyme systems with increasing early development

Early development of the African catfish Clarias gariepinus (Burchell, 1822), focusing on the ontogeny of selected organs

Embryonic development of Clarias gariepinus was studied from oocyte activation to the end of endogenous feeding (164 h post-fertilization, 164 h-PF). The ontogeny of the eyes, the ear, the heart, the digestive tract and the notochord were described histologically: (i) eyes were not pigmented at hatching (40 h-PF) and the anlage of the retina was observed at 96 h-PF. At 164 h-PF, the pigmented epithelium was distinguished as a single layer of cubical cells and the outer nuclear layer was composed of cones; (ii) the inner ear was oval-shaped and two otoliths and a sensory epithelium were differentiated at hatching. Three semicircular canals with rudimentary cristae developed until 96 h-PF, leading to a fully developed auditory vesicle at 120 h-PF; (iii) at 40 h-PF, a beating heart appeared as a thin-walled tube and at 72 h-PF no blood elements were recorded in the two-chambered heart. The four compartments (bulbus arteriosus, ventricle, atrium, sinus venosus) and mature blood elements were found only at 144 h-PF; (iv) the hatchlings possessed an undifferentiated digestive system, and differentiation of a buccal cavity, oesophagus and intestine was only recorded 96 h-PF. The connection between the oesophagus and the intestine and the anus opened at 144 h-PF; (v) the notochord in newly hatched embryos was composed of vacuolated cells and a peripheral layer of notochordal epithelium inside a fibrous sheath. Subsequently, vacuolated cells proliferated and notochordal epithelium changed from cubical to squamous cells. Eye differentiation was concomitant with first feeding, thus suggesting a key role of visual senses in feeding behaviour

Monitoring of DNA breakage in embryonic stages of the African catfish Clarias gariepinus (Burchell, 1822) after exposure to lead nitrate using alkaline comet assay

Increasing lead contamination in Egyptian ecosystems and high lead concentrations in food items have raised concern for human health and stimulated studies on monitoring ecotoxicological impact of lead-caused genotoxicity. In this work, the alkaline comet assay was modified for monitoring DNA strand breakage in sensitive early life stages of the African catfish Clarias gariepinus. Following exposure to 100, 300, and 500 microg/L lead nitrate, DNA strand breakage was quantified in embryos at 30, 48, 96, 144, and 168 h post-fertilization (PFS). For quantitative analysis, four commonly used parameters (tail % DNA, %TDNA; head % DNA, %HDNA; tail length, TL; tail moment, TM) were analyzed in 96 nuclei (in triplicates) at each sampling point. The parameter %TDNA revealed highest resolution and lowest variation. A strong correlation between lead concentration, time of exposure, and DNA strand breakage was observed. Here, genotoxicity detected by comet assay preceded the manifested malformations assessed with conventional histology. Qualitative evaluation was carried out using five categories are as follows: undamaged (%TDNA 75%) nuclei confirming a dose and time-dependent shift towards increased frequencies of highly and extremely damaged nuclei. A protective capacity provided by a hardened chorion is a an interesting finding in this study as DNA damage in the prehatching stages 30 h-PFS and 48 h-PFS was low in all treatments (qualitative and quantitative analyses). These results clearly show that the comet assay is a sensitive tool for the detection of genotoxicity in vulnerable early life stages of the African catfish and is a method more sensitive than histological parameters for monitoring genotoxic effects. 2008 Wiley Periodicals, Inc

The predominant role of 18F-FDG PET/CT over MDCT in assessment of ovarian cancer patient

Objectives: This article discusses that FDG PET/CT is superior to MDCT in evaluation of ovarian cancer oncological evaluation. Patients and methods: 87 PET/CT scans of 64 women with clinically suspected or pathologically proven ovarian cancer were retrospectively analyzed. The findings of contrast enhanced MDCT (CE-CT) were interpreted by two experienced radiologists unaware of PET/CT findings. At least two experienced nuclear medicine physicians who were unaware of CE-MDCT findings examined PET images, evaluating localization and characterization and comparing them to co-registered PET/CT images. Diagnostic accuracy was determined on a patient level and a region level. Results: PET has significantly higher Sensitivity, specificity, PPV, NPV, and overall accuracy of 94.7%, 86.7%, 93.1%, 89.7%, and 91.9% respectively compared to 89.5%, 30%, 70.8%, 60% and 68.9% for MDCT on patient level. The diagnostic performance of PET was also better at most anatomical sites when results were analyzed on region level. Conclusions and recommendations: FDG PET in addition to conventional imaging modalities should represent an important step in the diagnostic flowchart of ovarian cancer patients for evaluating abdominal and extra-abdominal probable metastatic deposits