صابر خدابنده

In recent years, many bioactive compounds has been extracted, described and purified from cephalopods. These invertebrates are a very rich source of sulfated polysaccharides with new structures.  The aim of this study was investigation of anticoagulant Glycosaminoglycans extracted from Sepia Pharaonis tentacles. Following the separation of their tentacles terminal portion, 21g of the wet weight was freeze-dried. First, glycosaminoglycans were extracted using 5g wet weight and 4g dry weight, separately, by Cetyl Piridinium Chloride cationic salt. In order to do identification of extracted glycosaminoglycans and compare with heparin, the FTIR spectrum was used. Activated Partial Thromboplastin Time (APTT) and the Prothrombin Time (PT) methods were used for anticoagulation properties. Results showed that 5g (wet weight) and 4 g (dry weight) of tentacles contain 250 and 800 mg of the Glycosaminoglycan, respectively. Also, the FTIR spectrum results confirmed the presence of heparin-like compositions among extracted glycosaminoglycans. Investigating the anticoagulation properties indicated that the extracted compositions have this property and increasing concentration enhance the clotting time of the human blood, significantly. According to the present study results, squid tentacles contain glycosaminoglycans which could act like heparin but significantly weaker antithrombotic feature than heparin.

Dunaliella salina microalgae is a unique eukaryotic microorganism that has a high range of tolerance against environmental fluctuations including salinity, temperature, and light. The aim of this study was to determine the growth factors and content of fatty acids in D. salina under salinity and ultraviolet conditions. The experiments were conducted at three different salinities (150, 200 and 250 ppt) under UVB and UVC irradiations in 3 replicates. Algae growth rate and total algae biomass were measured by counting the number of cells every alternate day for 11 days by hemacytometer. The fatty acids profile was measured by gas chromatography and the amount of total fat using a soxhlet device. The growth rate of algae in most of the treatments under high salinity stress, UVB and UVC irradiation was significantly lower compared to the control group (P≤0.05). The highest cell counts were detected in the control one (2.4 million cells/ml) and UVB-150 treatment (1.98 million cells/ml) on day 7. But despite the higher cell count in the control group, the highest fresh wet weight of algae was obtained in the UVC-200 ppt/salinity group. Substantial differences were detected in the fatty acids profile between the treatments (P≤0.05). Algae cultured under UVB-200 contained the highest PUFA-n3 (20.05%) and the sum of PUFA (32.05%), while the highest PUFA-n6 (13.96%) was found in the UVC-150 group. Algae cultured at 250 ppt salinity without UV irradiation had the highest level (52.75%) of saturated fatty acids (SFA). It was concluded that UV irradiations and high salinity suppresses the growth but improves the synthesis of polyunsaturated fatty acids.

Diabetes mellitus is a major health problem in the worldwide. Inhibition of DPP-IV is one of the methods to control diabetes type 2. Inhibition of this enzyme may improve glycemic control in diabetics by preventing the rapid breakdown and there by prolonging the physiological action of incretin hormones. Furthermore, improving the antioxidant system in diabetic patients can prevent the occurrence of secondary disease caused by oxidative stress. Therefore, the head of skipjack tuna was hydrolyzed with alcalase enzyme (1/5% of raw material weight) for 4 hours, in order to produce a product with antidiabetic and antioxidant activities. The DPP-IV inhibition activity, DPPH radical scavenging activity and reducing power of hydrolysate were measured. The results showed that the skipjack tuna head protein hydrolysate possess bioactive properties in a concentration dependent manner and increasing the protein concentration leads to a significant increase in bioactive properties of hydrolyzed product (p≤0.05). The IC50 of protein hydrolysate in DPP-IV inhibition and DPPH radical scavenging activities were obtained 1.016±0.02 mg/ml and 0.297±0.015 mg/ml, respectively. Also the reducing power of hydrolysate was 0.176±0.002 in 2.5 mg/ml protein concentration. Overall, according to the obtained results, it can be concluded that protein hydrolysate of skipjack tuna head possess high antioxidant and antidiabetic activities in vitro, and can be used as a food additive to enhance health level if additional research be conducted.

The freshwater shrimp, Macrobrachium nipponense is an invasive species which has recently been reported in Anzali Wetland, Iran. It exhibited good tolerance and adaption in this wetland ecosystem. This study examined certain aspects of feeding of M. nipponense in three habitats of this wetland. Shrimps were randomly sampled from April 2016 to March 2017. The stomach contents were obtained from 367 specimens ranging in length from 4.2 cm to 6.9 cm. The empty stomach index (VI) showed that this shrimp was a voracious (0 ≤ VI < 20) species in all seasons expect winter, when 99% of the specimens had empty stomachs. Fourteen dietary items were categorized in the three habitats of the wetland, with phytoplankton, mollusks and detritus forms being the dominant food items in the western, central and eastern habitats respectively. The feeding precedence index (FP) revealed that the most abundant portion of food was subsidiary one (50 ≥ FP ≥ 10) and the highest proportions of subsidiary food were phytoplankton (24.5%), gastropods (34%) and detritus (29.11%) in the western, central and eastern habitats, respectively. Omnivorous feeding is one of the reasons for the success, high tolerance and adaptation of M. nipponense in the Anzali Wetland ecosystem.

 The present study has done on 20 pieces L. vannamei shrimp, with average weight 5.53±0.02 g, in 500-liter tanks with three replicates during 8 weeks. At the end, the shrimp intestine were fixed in Bouin's solution for classic histology by Hematoxylin-Eosin (H & E) and green light (GL) and for immunolocalization of the Na+,K+-ATPase enzyme (sodium - potassium pump) by Immunohistochemistry methods. The results showed that the midgut epithelium was covered by simple columnar cells and nucleus position was in basal region of cells and Na+,K+-ATPase enzyme observed in basal region of cells. Evaluation of rectal pad large lobes deep infoldings and distance between the lobes were observed. Lumen was observed in the middle of rectal pad and apical lobes were evident and columnar cells in the marginal infoldings with basal nucleus were observed. Small appendage that called posterior diverticulum was located in primary and in upper region of rectal pad. This region was tubular and it cells with basal nucleus were observed. Rectum was located in distal region of hindgut. This region was a short muscular tube and lined with a simple columnar epithelium with low infoldings and apical nucleus in these cells were observed. The Na+, K+-ATPase enzyme (sodium - potassium pump) was located in baso-latral area in all regions of hindgut cells. We concluded that the use of H&E and GL is an appropriate method to separate different parts of intestine in vannamei shrimp and also immunohistochemistry is a suitable method for Na+,K+-ATPase enzyme localization.

The present study investigated the inhibitory effects of the oral disk extract of sea anemone, Stichodactlya haddoni on acetylcholinesterase and caspase activity. The sea anemone samples were obtained from the intertidal zone of the Hormuz Island in the Persian Gulf. Millipore ultrafiltration was carried out to separate different fractions of the crude extract. Ellman method was used to measure acetylcholinesterase inhibition. The activity of caspase 3 was measured using a calorimetric assay kit of Caspase in breast cancer cell MCF-7. At the end of the experiment, the highest percentage of inhibition of the acetylcholinesterase was observed in 6 mg/ml of the 30-100 kDa fraction (p<0.05). The activity of caspase 3 in MCF-7 treated with concentrations of 400, 600 and 800 μg/ml from the fraction 100-30 kDa were significantly different after 12, 24 and 48 hours (p<0.05). The concentration of 600 μg/ml in the desired fraction showed the highest caspase activity after all the three mentioned times. In conclusion, the protein toxins of the sea anemone, S. haddoni, with a molecular weight of 30 to 100 kD can have anti-Alzheimer's effects by inhibiting acetylcholinesterase enzyme and the lethality effects on the breast cancer cell by activating caspase 3 and possibly inducing apoptosis.

The kidney of the mouse is a good model for the study of the structure and physiology of nephron in mammals, including humans. Between the laboratories animals, Nude mouse is used frequently as a model in the study of cancer. Due the importance of the relationship between sodium-potassium pump and cancer, the nephron structure and Immunolocalization of Na, K+ATPase in different parts of the kidney were examined in Nude mouse. Six Nude mouse from Pasteur Institute (Amol Branch) were prepared and their kidneys were isolated, then based on the usual histological method, dehydrating and embedding in paraffin were carried out. A number of tissue sections stained by using hematoxylin and eosin, and localization of Na, K+ATPase was performed using immunohistochemistry method. Histological observation showed that the kidney of the Nude mice composed of two parts: cortex and medulla, the cortex is the analogue of the human kidney cortex and the medulla is the analogue of the human kidney medulla. Immunohistochemistry photographs showed that the presence of Na, K+ATPas, as an indicator of the presence of sodium-potassium pump, is different in segments of kidney. The results of this study suggests that the difference of Na, K+ATPase intensity in different segments of kidney shows the different function of ions absorption in these segments. Also immunohistochemical method is useful for the changes of Na, K+ATPase presence, due to using of drugs and their side effects on the kidney function.

In the present study, the effects of enrichment of Artemia franciscana with fish oil and soybean oil accompanied with vitamin E were investigated on the structure of pyloric caeca in Persian sturgeon (Acipenser persicus) larvae. A total of 1500 Persian sturgeon larvae (with the average weight of 51±2 mg) were divided into 15 tanks. Larvae were fed for 17 days in 5 experimental treatments including the treatment C (non-enriched artemia), S15 (enriched artemia with soybean oil and 15% vitamin E), S30 (enriched artemia with soybean oil and 30% vitamin E), F15 (enriched artemia with fish oil and 15% vitamin E) and F30 (enriched artemia with fish oil and 30% vitamin E). From each traetment, 15 larvae were collected and were fixed in Bouin’s solution for histological studies. Histological results showed that both fish oil and soybean oil were the main and effective factors on the total area and histo area indices of the pyloric caeca and the enrichment of artemia with fish oil and soybean oil was significantly increased this factor as compared to that of the control treatment (P

Aims: Skipjack tuna has the highest level of catch rate among tuna all over the world. Its head contains about 64% protein. Many Protein Hydrolysates and peptides obtained from various marine sources have a high antioxidant power. The aim of this study was to investigate the antioxidant activity of Protein Hydrolysate in Skipjack tuna head.
In this experimental study, 30 Skipjack tunas were investigated. At first, the amount of different compounds (protein, fat, ash, and moisture) was evaluated in the raw material; then, the hydrolysis process was performed by Alcalase enzyme and the hydrolysis degree of the protein hydrolysate was evaluated at different times. The antioxidant activity of the protein hydrolysate mixture was measured by DPPH radical scavenging activity, iron revival power, and ABTS radical inhibitory activity. For data analysis, the analytical tests were used.
Findings: The main part of the fish head was protein and it had high levels of ash. The degree of hydrolysis increased with increasing time and was it significant at 15, 60, and 120 minutes (p Protein hydrolysate in Skipjack tuna head has a high antioxidant activity and can be used in food products to increase oxidation stability.

Background and Biological studies on marine fauna, especially invertebrates, has significantly increased in recent years which led to the identification of many different bioactive compounds. The sea cucumber are echinoderms with a very muscular body wall that contains 70% collagen and is considered a rich source of protein. Based on recent researches on bioactive compounds extracted from sea cucumber, it was found to have cytotoxic, antioxidant, anti-tumor, and anti-coagulation properties.
In this experimental study, enzymatic hydrolysis method was used to study the anticoagulant properties of hydrolysates protein in muscles of sea cucumber. Finally, the anticoagulant properties of hydrolysates protein on the human blood plasma was examined by the Activated Partial Thromboplastin Time anticoagulant test (APTT) in two concentrations (90 and 130 µg/ml), and Prothrombin Time (PT) in different concentrations (220, 440, 670, and 900 µg/ml).
The total amount of hydrolysates protein was found to be 55.8 mg/g in wet tissue. The results of anti-coagulation assays showed that the hydrolysates protein of the sea cucumber muscle contains anticoagulant properties on human blood plasma and could prolong the clotting time.
Peptides from the hydrolysis in sea cucumber muscle have anticoagulant properties as already reported for heparin-like compounds.

In this study, the effect of abrupt transfer from freshwater (FW) to the Caspian Sea Water (CSW, 11‰) on ion-regulation capacity of intestine spiral valve were investigated in juvenile Persian sturgeon Acipenser persicus (2.55±0.41g). Immunofluorescence localization of ion transporter proteins including Na,K+ATPase (NKA), Na,K,2Cl– (NKCC) and cystic fibrosis transmembrane conductance regulator (CFTR) were performed 10 days after abrupt transfer. Co-localization of NKA and NKCC in the cytoplasm of enterocytes for both FW- and CSW-acclimated fish indicated that these two proteins were not affected by salinity transfer. Furthermore, CFTR was also found in the cytoplasm but mostly in apical membrane of spiral intestine enterocytes. Based on the obtained results, the main function of spiral valve is nutrient absorption and indirectly has a minor role in ion regulation in juvenile Persian sturgeon. Furthermore, this organ has good physiological development condition in 2-3g fish and it prepares their ability to expose to the Caspian Sea water salinity.

Protein compounds were extracted from the mucus of sea anemone, Stichodactyla haddoni, and their effects on the gills of rainbow trout, Oncorhynchus mykisswere examined. Sea anemone samples were collected from the intertidal zone of the eastern coast of Hormuz Islandand frozen samples were transported to the laboratory. Then the mucus was extracted using of the PBS solvent and doses of 5, 10 and 24 mg/dry weight of total protein was injected into the tail vein of the fish. Upon the inactivation of fish, histopathological changes were examined using of the classical histological method. Lethal signs were observed in the gills, including aneurysm, hypertrophy of epithelial cells, lamella clubbing and deformation, subepithelial edema, lamella congestion in the interlamellar region and necrosis. The damages were more serious with increasing doses. The results showed that protein compositions of the mucus can cause numerous lesions in the gill tissue of fish, which act as an excretory, respiratory and ionic regulation tissue, the failure of which can lead to failure of fish’s vital functions that can be one of the reasons for the death of the hunted fish.
The results showed that the protein compositions of mucus can cause numerous lesions in the gill tissue, as an excretory, respiratory and ionic regulation tissue lead to failure of it functions that itself can be one of the reasons hunted fish death.

Background and The 4T1 cell line is a laboratory model used in the study of tumors biology. This cell line is very tumorigenic with high metastatic capacity in different organs. In this study, histology and immunohistochemistry methods were used to investigate the structure and localization of Naﲯ ATPase enzyme in 4T1 cells induced breast cancer tumor in Balb/c nu mice.
The histological sections (4 µm) were stained using Hematoxilin-Eosin and IgGα5 special antibody was used for immunohistochemistry study.
Study of tumor structure showed abnormal proliferation and high mitogenicity in epithelial cells with high proliferation rate. Immunohistochemistry analysis revealed significant immunofluorescence, indicating abundant presence of NaﲯATPase enzymes (sodium potassium pump) in tumor cells.
Current results showed high rate of sodium potassium pump in plasma membrane of tumor cells. Immunofluorescence characteristic of 4T1 tumor cell lines make them appropriate candidate for antitumor studies.

The response surface methodology was employed to optimize the effects of pH, temperature (˚C), time (min) and the ratio of enzyme to substrate (% of substrate) on the hydrolysis process of cuttlefish muscle by alcalase. Central composite rotatable design with 5 levels and 4 factors and α=2 was used for the optimization of the process to gain the highest degree of hydrolysis. pH, temperature, time, enzyme concentration, interaction of temperature-enzyme concentration, square of pH, temperature, time and enzyme concentration had significant effects on the process. The R2 = 0.95, lack of fit

Sea anemone is ocean dwelling and typically organisms sessile. These tentacles produce a variety of peptides and proteins that act as, neurotoxins and cytolysins toxins. Cytolysins due to their effect on specific tissue and lysing properties are known as antiparasitic or antitumor compounds.
Sea anemone Stichodactyla haddoni were collected from the coast of Hormuz Island in November 2012 and tentacles extraction was performed by using of saline PBS solvents. Peptides above 10kDa and peptides below 10kDa were separated by amicon® Ultra-15 10K device. The hemolytic activity was assessed by the micro hemolytic method for human, rainbow trout Oncorhynchus mykiss and common carp Cyprinus carpio erythrocytes.
Uniform red color suspension in the wells considered as positive hemolysis. The crude extracts, peptides above 10kDa and peptides below 10kDa at various concentrations were showed hemolytic effect on human, common carp Cyprinus carpioand rainbow trout Oncorhynchus mykiss erythrocytes.
The crude extract, peptides above 10kDa and peptides below 10kDa have a hemolytic effect on human erythrocytes and two species of fish. The venoms showed the most hemolytic activities on human blood among others.

In this study, the combined effect of different levels of dietary n-3 HUFAs with α-Tocopherol on changes in the structure of intestine and pyloric caeca and fatty acids profile of Caspian salmon, Salmotrutta caspius, fry (initial weight of 600 ± 25 mg for 10 weeks) were investigated. Six experimental diets containing three different dietary levels of n-3 HUFAs (Low: 1 0.5, DHA EPA, Medium: 2 1, DHA EPA, High: 4 2, DHA EPA g/100g diet) with two different levels of α-Tocopherol (Low: 300 and High: 1000 mg/kg diet) were prepared and named: LL, LH, ML, MH, HL, HH (HUFA/α-Tocopherol), respectively. The results showed that increase in dietary vitamin E increased HUFA levels in polar lipids, particularly when dietary HUFA were higher. Histological analysis showed the mean length of enterocytes had not significant difference in anterior intestine and pyloric caeca segments. However, in fish fed moderate and high HUFA diets enterocytes were slightly higher. Feeding Caspian salmon fry with moderate and high levels of omega 3 HUFA increased the villi area of mucus layer in anterior intestinal. The results of this study showed a concomitant increase in dietary omega 3 HUFA and vitamin E increased long chain fatty acids, especially in moderate and high levels of HUFA fatty acids, which these fatty acids have beneficial effects on the growth and health of fish.

A high population of the sea anemone inhabit in the intertidal zone of the Hormoz Island. The tidal zones generally are characterized by severe environment stresses (vis, UV and high temperature), that can increase the intracellular oxidation and free radicals as well. Oxidation and free radicals is paved by the animal defensive mechanisms, including accumulation of antioxidant compounds and enzymes. In this study, carpet anemones sampled from intertidal zone of the eastern city of Hormoz Island (IRAN). Extraction was performed by using of PBS and methanol 40% solvents: 2 g dry weight of the mucous, by 2 replications, and 10 g wet weight of the oral plate, by 6 replications, from 6 samples. Different concentrations of extracts were made for antioxidant tests. The antioxidant properties of extract were measured by DPPH and FRC methods. Results showed that a correlation is between the antioxidant activity and the concentration of extracts in two methods. In DPPH assay, IC50 values was 1.469±0.208, 1.85±0.016 for mucus extract by PBS and methanol 40%, respectively, and it was 0.733±0.06, 0.444±0.036 mg/ml, for oral plate extract by PBS and methanol 40%. Our results showed that the antioxidant activity of oral plate was significantly higher than mucous in this anemone.

The effects of mucal proteins of sea anemone, Stichodactylahaddoni,on different stages of embryonic development of zebra fish (D. rerio) were examined. The sea anemone samples were collected from the intertidal areas of the Hormuz Island (Persian Gulf), and were frozen at -160 °C. Protein and peptide components were extracted by 100% methanol. Following the total protein assessment by ELISA, three concentrations (2.1, 3.7 and 7.4 mg/ml distil water) were prepared. From each concentration, 2 ml was added to the microplates containing 150 zebra fish eggs each, with 2 replications; microplates with normal aquarium water was also used as control group. The eggs were incubated for 72 hrs and the process of embryonic development was observed every 6 to 12 hours. Results showed that the embryonic development was normal in the control group, while the eggs treated with 3.7 and 7.4 mg/ml ofmucal proteins degenerated and blackened in less than 12 hours. Also a delay in the phase of growth in embryonic development was observed in the group with 1.2 mg/ml of protein. Our results showed that the mucal proteins from this sea anemone can affect embryonic development rapidly, causing delayed growth at low concentration, and cell lysis and embryonic degeneration at high concentrations.

The marine environment is anexceptional reservoir of bioactive natural products, many of them exhibit structural/chemical features that not found in terrestrial natural products.Glycosaminoglycans are one of this various bioactive compounds. Heparin, as a well known glycosaminoglycan, is a sulfated glycosaminoglycan that has natural anticoagulant properties. Heparin and heparin-like compounds are used as anticoagulants in many aspects of medicine. However, for two main reasons: 1. Contamination in heparin samples obtained from pig intestine or bovine lung pathogens and other pathogens, 2. resource for use of heparin is limited and there are a lot of requirements for new compounds from natural resources. According to GAGs importance and widespread using of heparin in medicine, in the present study, GAGs compounds extracted from sea anemones and anticoagulant properties of the human blood is investigated. GAGs compound was extracted by using cetylpyridinium chloride. Anticoagulation activity of extracted GAGs (the extracted tentacle) was tested in human blood plasma, using manual procedures, and assay system, prothrombin time (PT) and activated partial thromboplastin time (APTT). In this study the amount of the crude GAGs was 24 mg per gram of tentacle dry weight. The results ofanticoagulant activity extracted on human blood plasma showed that these compounds prolonged clotting time compared to the control. In APTT and PT assay of the extracted GAGs from the sea anemone also clotting time prolonged in compared to the control. The results demonstrated that anticoagulant compounds existed in the tentacle of the sea anemone, and although their effects is weaker than the heparin, but they can be substituted for heparin, at least in laboratory conditions.

The lethal concentration and nephrotoxic effects of sea anemone, Stichodactyla sp., mucus on zebra fish, Danio rerio were examined. Sea carpet was collected from an intertidal zone of the Hormoz Island, frozen in nitrogen tank (-160°C), and transported to the laboratory to have its mucus isolated. The lethal concentration (LC5096h) of mucus for the fish, determined as 175.03 mg/L. Three concentrations of mucus (250, 500, 1000 mg/L) were made and added to the fish aquaria. Then, the histological changes of the fish kidney were examined. In 1000 mg/L, the fish was killed in less than 2h and pathologic lesions of the kidney were evident, including congestion, enlargement and necrosis of the glomerulus, reduction of Bowman’s space, fluids inside tubular and glomerular capillaries inflation in interstitial space, but in 500 and 250 mg/L, no death were observed and a slight increase in glomerular volume was observed. Results showed that mucus of the sea anemone could be harmful and even lethal to the zebra fish by damaging its kidney, the severity of which depended on its concentration.

To determination the toxic effects of atrazine on Caspian kutum, the 96-h LC50 of atrazine was measured for newly hatched larvae as 18.53 ppm. Toxicity of atrazine herbicide on Caspian kutum larvae was investigated using concentrations: 9.25ppm, 4.62 ppm and 2.31 ppm for 7 days. Comparison of the length, weight and condition factor showed that no significant differences between atrazine exposed and control groups. The concentration of Na+, K+, Ca2+, Mg2+ and Cl- in whole body of larvae in control and atrazine exposure groups were measured and the results showed that concentrations of all these ions is higher in atrazine exposure group than control group. Results of the present study showed that atrazine could affect the ion compositions of the body even at sublethal concentration and acute exposure but at this kind of condition have no effects on growth parameters of the body.

Cone snails (genus: Conus) are groups of marine mollusks that make a group of toxin compounds (conotoxins) for different purposes. Several studies proved the medical properties of conotoxins and existence of different toxic compounds in different parts of venom apparatus. So, recognizing the simplest and the most suitable method for separating the venom apparatus and purification of conotoxins is necessary. For this purpose, several specimens of Conus terebra thomasi were collected from the Northern sandy coast of Larak Island and the shells of samples were cracked by vice. After pulling out the snail from its shell, drawing out of the venom apparatus was done by cutting the posterior section of foot muscle. Venom apparatus was divided into three parts, venom bulb, proximal and distal venom duct and each sample was freeze-dried. Extraction was done by three solvents (Acetone 100%, A mixture of Acetone100%-Methanol20% and Phosphate Buffered Saline) and the total protein of each extract was determined by Bradford protein assay. The results showed that the amount of total protein in two extracts (Acetone and Acetone-Methanol) were very low and cannot be measured by this method, but it is measurable in third extract (venom bulb: 38.79, proximal venom duct: 52.4 and distal venom duct: 46.01 mg/g dried tissue). By using electrophoresis method, different conopeptides were shown in different parts of venom apparatus. Also, the most suitable solvent between the checked solvents in this study is phosphate buffered saline because of high concentration of extracted conopeptides, and reducing the risk of tissue damage. Meanwhile, the extracted compounds are suitable for investigating their bioactivity.

In this study¡ the extraction of peptides from sea anemone Stichodactyla haddoni mucus andtheir effects on the gills of rainbow trout Oncorhynchus mykisswere examined. For this purpose¡ Specimens of S. haddoni were collected from the intertidal coast from Hormuz Island. Frozen samples were transported to the laboratory.Then the mucus was extracted using of the PBS solvent.Peptides down of 10kDa were separated by amicon® Ultra-15 10K millipore filter device. These peptides were injected into the caudal vein of the fish and after inactived¡ histopathological changes were examined using of the classical histological and immunohistochemical method and also plasma osmolarity and ions were examined in this fish. The various tissue damages of the gill were observed including hypertrophy¡ lamella deformation¡ subepithelial edema¡ lamella congestion in the interlamellar region and necrosis. The injection of these peptides also reduced blood plasma ions and osmolarity. The results showed that the extracted peptides from sea anemone mucus can cause numerous anomalies in the gill tissue¡ change in blood plasma ions and osmolarity of rainbow trout and finally resulted in failure of physiological osmoregulation function of the gills and finally led to the fish death.

To investigate the branchial structure of the Caspian Kutum we used releasing fry’s (21-33 mg Body Weight). Study of the branchial tissue showed that in these fry’s branchial system was made up of 4 gill arch in each side of the head، one pseudobranch in each side of the head and one hemibranch at the posterior wall of the branchial chamber in each side of the head. Each gill arch bears gill filaments and lamella. Pseudobranch tissue is very similar to main gill tissue but it’s made up of lamellar shaped structures bound to each other. Immunolocalization of the chloride cells showed that these cells were located on the gill arch، gill filament، at the basis of the lamellae، pseudobranch and the hemibranch of the posterior wall of the gill chamber. Measurements of these cells showed that mean length was 9. 70±0. 28 µm and mean width was 6. 83±0. 24 µm. Ultrastructural studies showed that the most surface of the gill epithelia was composed of the pavement cells that have apical microridges and between them one can see the chloride cells with apical pit.

The combined effect of different levels of dietary n-3 HUFAs [eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA)] with α-Tocopherol on growth parameters (final weight, SGR, FCR, and some health indices (hematocrit, plasma lipid and lipoprotein levels) of the Caspian trout fry, Salmotruttacaspius, was investigated. Six experimental diets containing three different dietary levels of n-3 HUFAs (Low: 1 0.5, DHA EPA, Medium: 2 1, DHA EPA, High: 4 2, DHA EPA g/100g diet) with two different levels of α-Tocopherol (Low: 300 and High: 1000 mg/kg diet) were prepared and named: LL, LH, ML, MH, HL, HH (HUFA/α-Tocopherol) groups, respectively. Fries (600 ± 25 mg) were randomly distributed in tanks (50 fries per tank) and were fed with the experimental diets for 10 weeks. The work was conducted in triplicates. Results showed that the SGR and body weight were significantly higher in the MH group than other groups. Hematological parameters, especially plasma lipids and lipoproteins were more influenced by the dietary n-3 HUFA and vitamin E and interaction between vitamin E and n-3 HUFA has minimal effect on these parameters. Results of this study suggested that increasing of dietary n-3 HUFA could improved growth performance and enhance health in the Caspian trout fry when appropriate level of α- Tocopherol supplemented.