الفهرس 
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Abstract
The present work dealt with the study of some species of Decapodiformes (squids and cuttlefishes) collected from the Egyptian coast from the Red Sea which were identified by using the identification keys.
Squid Sepioteuthis lessoniana were identified according to FAO species catalogue Vol. 2, Cuttlefishes Sepia pharaonis and Sepia orbignyana were identified according to FAO species catalogue Vol. 1 and Identification guide for cephalopod.
The present study also include, in details, the ink apparatus and the funnel organ of the squid Sepioteuthis lessoniana as the secreted ink of cephalopods is a mixture of two glandular secretions: ink gland which produces a black ink containing melanin and the funnel organ (a mucus-producing gland) which located on the internal surface of the funnel.
Histological and histochemical techniques used to study the structure of this apparatus and the funnel organ. The ink apparatus composed of ink sac, ink gland and ink duct. The ink sac consists of four layers: outer connective tissue capsule, muscularis, sub-mucosa and mucosa. Ink gland lie in the cavity of the ink sac and consists of numerous perforated lamella of connective tissue covered with one layer of glandular epithelium and differentiated into two different regions: the caudal region (formative zone) and the apical region.
By using Ferro-ferricyanide method for the detection of melanin granules, the caudal region of the gland, differentiated into two zones: an inner smaller zone with immature cells that lack melanin granules and larger outer zone that consists of mature melanin producing cells and the cavity of the gland in this caudal region contains no or little amount of ink. However, in the apical region, all the epithelial cells are mature melanin producing cells with very little amount of melanin granules at its apex and the cavity appeared with large amount of ink.
The funnel organ of both squids and cuttlefishes understudy consists of two lateral pads and a median pad. Both the lateral and median pads consist of epithelial and sub-epithelial layer. The epithelial layer composed of single layer of columnar cells while, the sub-epithelial layer contains many secretory cells that may be aggregated in clusters or may be solitary.
The present study also includes screening some of the medical applications of the ink of the cuttlefishe Sepia pharaonis.
Antibacterial and antifungal activity of the ink were tested by using the ink in its liquid form, lyophilized powder of the ink and the lyophilized powder of the ink gland.
The ink of Sepia pharaonis was dissolved in dist.water, acetone, ethanol, methanol, saline solution and DMSO then tested on 6 bacterial strains (Bacillus cereus, Escherichia coli, Micrococcus luteus, Pseudomonas aeruginosa, Serratia marcescens and Staphylococcus aureus) and 6 fungal strains (Aspergillus flavus, Candida albicans, Fusarium oxysporum, Geotrichum candidum, Scopulariopsis brevicaulis and Trichophyton rubrum), however the results revealed that these extracts didn’t exhibit any antimicrobial activities against tested bacteria or fungi except in the case of methanol extracts of the ink and ink gland which tested against Staphylococcus aureus as they exhibit trace activity with inhibition zones: 6mm- 8mm.
Seawater and Tris-HCL also were used as solvents (with ultrasonication) for the ink of Sepia pharaonis and tested on Escherichia coli, Staphylococcus aureus, Aspergillus flavus and Candida albicans. The results revealed that the ink with these solvents also didn’t exhibit any antimicrobial activity.
Cytotoxic activity of the ink of Sepia pharaonis was also tested as a medical usage of the ink. The defatted lyophilized powder of the ink was tested against two cancer cell line, MDA-MB-231 breast cancer cells and Hep G2 hepatocellular carcinoma cells after 24 hrs of treatment with different ink concentrations (0.0, 1, 10, 100 and 1000 µg/ ml ) using MTT assay (cell viability assay).
The results showed that the viability of the cells decreases with the increased concentration of the ink in both types of cancer cells. In the case of breast cancer cells MDA-MB-231, cell viability was found to be 83.7% with a concentration of 1μg / ml; 78.94% with concentration (10μg / ml); 76.9% with concentration (100μg / ml); 30.9% with concentration (1000μg / ml). The highest toxicity of cells (69%) was recorded with the concentration of (1000μg / m) and so the ink had a significant toxic effect (P <0.001) with all concentrations that used and IC50 was recorded as (651.9 μg / ml).
In the case of Hep G2 cells, the results also showed that the ink had a toxic effect on these cells. The highest toxicity of the cells was recorded as 45% at the concentration 1000 μg / ml and the lowest percentage of cell toxicity was 20% with the concentration 1 μg /ml. IC50 was recoreded as 1156 μg / mL. Thus, the ink had a significant cytotoxic effect (P <0.05) at the concentration of (1 μg / ml) and P <0.01) at the concentrations (10, 100 μg / ml) and P <0.001 at 1000 μg /ml).
The present study also includes the morphological characterization of hemocytes of the cuttlefish Sepia pharaonis and the histological structure of its hemopoietic organ (white body).
Haemolymph was collected from the branchial heart then blood smears were prepared and stained with (H&E). Results revealed that there are three main types of hemocytes in the haemolymph of Sepia pharaonis: hyalinoytes, granulocytes and agranulocytes. The granulocytes divided into two sub-types: a) granulocytes with acidophilic granules and cytoplasm, b) granulocytes with acidophilic granules and basophilic cytoplasm. Agranulocytes also can be divided into two sub-types: a) Agranulocytes with acidophilic cytoplasm and b) Agranulocytes with basophilic cytoplasm.
The histological structure of the hemopoietic organ (white body) of Sepia pharaonis indicated that it is a multilobed organ enclosed by a thin fibrous connective tissue capsule that consists of collagenous fibers. Hemocytes within the white body were enclosed in a supportive connective tissue fibers that form a network of small unites in which hemocytes are grouped in clusters.