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Histopathological and pathomorphological effects of 15 ppb mercuric chloride on Persian sturgeon, Acipenser persicus, were investigated using histological and electron microscopy observations. Light microscopy showed that the gill epithelial hypertrophy, wrinkling and hyperplasia in lamellar epithelia and lamellae fusion occurred after 48 h of exposure. Gill epithelia also showed occasional necrosis, which had almost been completed and blood emerged from the capillaries. However, occasional necrosis in some regions of the filament, both with blood emerging and with no bleeding, was observed by using electron microscopy. These injuries were well observed in inter-lamellar regions of the filament and also wrinkling of the lamellar epithelium. Ultrastructural observations showed some cellular disorders in gill epithelium of the Persian sturgeon, A. persicus, fry. In addition, increase in apical vesicles of the chloride cells and necrosis in apical surfaces of some chloride cells, hypertrophy and necrosis of the chloride cells’ mitochondrion and endoplasmic reticulum also were some of the other cellular disorders observed through transmission electron microscopy. In conclusion, the gills of A. persicus fry were sensitive to low concentrations of inorganic mercury (HgCl2).

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The effects of fish weight on salinity tolerance were studied in Caspian salmon (Salmo trutta caspius) parr. 180 fish (all with 2 years old but with three weights; 5, 15, 25g) were selected and they reared in freshwater (FW) and brackish water (BW; 13ppt salinity) for 10 days. The mRNA expression of two α-subunit isoforms of Na⁺, K⁺-ATPase (α1a and α1b) and NKCC co-transporters were studied in their gill tissue. In all three weight groups, the mRNA levels for the α1a isoforms decreased following BW exposure, whereas α1b levels significantly increased in 15g and 25g groups. In addition, NKCC gene expression were significantly higher in the groups of BW than FW in 15g and 25g weights (P<0.05). The reciprocal expression of Na⁺, K⁺-ATPase isoforms (α1a and α1b) during salinity acclimation suggests that they may have different roles in the gill of FW and BW fishes; ion uptake in FW and ion secretion in BW. In conclusion, in the Caspian salmon, between parrs with the same age, the group with the weight of 15g possesses better compatibility with BW than to other groups.  After reaching to 25g, fish passed smoltification and they became more compatible with the FW environment and maybe lost its osmoregulation ability in saline or brackish water.

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The gill Structure and localization of Na⁺, K⁺-ATPase were examined through branchial arches of the Persian sturgeon, Acipenser persicus larvae (25 days-post-hatched, 417.3 mg). Studies were conducted through light microscopy (H&E Staining) and immunofluorescence for Na⁺, K⁺-ATPase. Results showed each gill consisted of four complete holobranches and opercular hemibranch. Each filament carried rows of lamellae consisting of a network of interconnecting blood lacunae, which were lined by pillar cell flanges. Pavement cells covered the outermost layer of the lamella and blood cells were found in lacunae. High density of ionocytes (529.73 per mm² of the gill tissue) was found at the base of the lamella, in the interlamellar regions, on the filaments and the septums. Ionocytes possessed large size and round basal nuclei. Ionocytes possessed strong immunofleurescence in their cytoplasm, especially in the basolateral sides because of high concentration of the enzyme. The results showed that the main structures of the gill has already been formed at this developmental stage of the Persian sturgeon, and along with its respiratory and excretory roles, it also plays an important role in osmoregulation.

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Cone snails of the genus Conus are highly regarded for their medicinal compounds derived from their toxins. In order to examine the venom apparatus structure, 12 specimens of C. textile were collected from the coastal zone of Gheshm Island and fixed in Bouin's for 48 hours and transferred to laboratory in ethanol. After breaking of shellfish, the venom apparatus were isolated and their different parts (after molding and cutting) were stained by HE and HEG and photographed by Nikon microscope. The stereomicroscope observation showed that the venom apparatus consisted of  (1) toxin production part (venom duct), (2) toxin transmission part (venom bulb), and (3) injection part (radula and proposcis). Photographs of sections showed that the venom bulb was completely muscular, consisting of longitudinal and transverse muscle fibers, and in their middle part a channel with epithelial cells was observed. Venom duct walls composed of 3 parts including the outer layer of muscle an inner layer of columnar epithelial cells with basal nucleus and the inner lumens which filled by the granules. HEG stained slides showed a much sharper cytoplasmic and nuclear implementation, particularly granules containing toxins were easily countable and measurable. Although the onventional HE staining method clearly showed different parts of the gland, but HEG method in addition to distinguishing different sections of tissue, seemed to be a suitable technique for studying the role of different parts of the organ in producing conotoxin in the form of secretory granules.

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Morphological changes of the chloride cells and the α_1b subunit gene expression of Na⁺-K⁺-ATPase in triploid rainbow trout (70.6 g average weight) were studied upon direct transferring to 6, 12 and 18 ppt salinities. Changes in abundance, distribution pattern, and the sectioned area of the chloride cells was studied through classic histology and Na⁺ K⁺-ATPase localization was performed through immunofluorescence light microscopy using a mouse monoclonal antibody IgGα5. Gene expression of Na⁺-K⁺-ATPase α_1b subunit was studied by semi-quantitative gene expression methods.No mortality occurred among the fish in all salinities during the 10-days experimental period and treated fish kept their plasma osmolality at standard physiologic levels. All the fish also showed similar distribution pattern in their chloride cells that were distributed on filaments, between and over lamella. Histological studies confirmed some abnormal morphological changes such as lamella interruption. Immunohistochemical studies showed the highest number of the chloride cells on lamella and between lamella in 18 ppt and the maximum sectional area of the chloride cells in freshwater. Gene expression of Na⁺-K⁺-ATPase α_1b subunit had direct correlation with increasing trend of salinity. In conclusions, triploid rainbow trout was found to be adaptable to the various experimented salinities and could be recommended for rearing in brackish water.

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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.

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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.

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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 < 0.05 and adeq-Precision of 14.16 for the model showed that the model could explain the variability within the range of values. The optimum condition for 42.0117 % of degree of hydrolysis was determined by Design Expert as pH 8.19, temperature 50.23, time 129.62 and enzyme2.15%.

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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.

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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.
Materials & Methods: 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<0.05), but not significant at 120 and 240 minutes (p<0.05). DPPH radical scavenging activity increased with increasing hydrolysis time and there was a significant difference in all samples obtained from different times (p<0.05). The iron reduction capacity of the protein hydrolysate samples increased with increasing the hydrolysis time, and the highest amount was at 240 minute. The samples obtained from different times had a significant difference in iron reduction capacity (p<0.05). Increasing the concentration of protein hydrolysate increased inhibitory activity (p<0.05).
Conclusion: Protein hydrolysate in Skipjack tuna head has a high antioxidant activity and can be used in food products to increase oxidation stability.

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Abstract 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.

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The ultrastructure of the cells, Na+, K+-ATPase activity and immunolocalization were examined in the barnchial chamber of Libellula lydia (Drury, 1773) larvae. Na+,K+-ATPase activity and localization were performed through biochemical techniques and immunofluorescence light microscopy using a mouse monoclonal antibody IgGα5, respectively. The branchial chamber possesses six pair gills lamellae that extend into the rectal lumen. A thickened epithelial layer and a modified fat body cells layer are present at the base of the each gill lamella. Epithelial cells covered by a thin cuticle and they possess apical microvilli and baso-lateral membrane infoldings associated with mitochondria. The cytoplasm of the modified fat body cells is filled with mitochondria, glycogen and a few lipid droplets. The Na+,K+-ATPase activity was significantly higher (15.36 µM Pi mg-1 protein h-1) in the branchial chamber. Na+,K+-ATPase immunofluorescence staining was observed in the epithelial layer cells of the basal pads of the rectal gill lamellae, with a consistently high immunoreactivity. These findings show that the epithelial cells present cytological features of the ionocytes, a high activity and concentration of Na+,K+-ATPase, confirming their participation in osmoregulation through active ion exchanges.

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The tadpole shrimp of genus Triops is a well-known living fossil whose fundamental morphology has been unchanged for 220 million years. We collected specimens of Triops cancriformis in temporary water bodies near the southern part of Urmia Lake (in the Fall of 2005). Some biological characteristics of this Triops were investigated. The feeding re-gime of T. cancriformis was found to be related to the fauna and flora of the temporary pools. Invertebrates and animal detritus were found to constitute major part of the feed-ing regime. The existence of Triops cysts and particles in the gut also showed certain de-gree of cannibalism. Morphological and histological investigations showed that the popu-lation of T. cancriformis was female and there was only one male among 400 samples col-lected. Observation of sperm among follicle ducts of a few samples indicated some degree of hermaphrodity, but the animal seemed to reproduce mainly through parthenogenesis. Fecundity, varying from 100 to 2500 cysts, was with a few exceptions related to the body size. The average cyst diameter was 40085 m.

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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.

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Aim: In this study, the antioxidant properties of hydrolyzed protein from longtail tuna dark muscle with commercial enzymes (alcalase, alkaline protease, and evatase) were investigated.
Materials & Methods: Protein hydrolysates from tuna dark muscle were prepared by different enzymes Degree of hydrolysis (DH) was performed by TCA technique. The five aliquots at 60, 180, 240, 300, and 360 min were gathered during hydrolysis. The antioxidant activity of aliquots was monitored by in vitro assays (DPPH inhibition ability and Ferric (Fe3+) reducing power).
Findings: The antioxidant activities of protein hydrolysate from tuna dark muscle (TDM) increase with increasing time and DH. Alcalase hydrolyzed protein (AHP) generally showed higher antioxidative activity than evatase hydrolyzed protein (EHP) and alkaline protease hydrolyzed protein (APHP). Among the samples (concentration 3 mg.ml^-1), AHP at 360 min significantly exhibited the highest ability to scavenge DPPH radical (72.6 %). Furthermore, AHP and APHP significantly showed a minimum IC50 value of 1.1 mg.ml^-1 at 240 and 360 min hydrolysis. APHP significantly exhibited the highest ferric reducing power of 0.83 at 300 min and 0.76 at 240 min. AHP and APHP significantly showed the highest ferric reducing power of 0.74 at 360 min (p < 0.05).
Conclusion: This study confirmed that protein hydrolysate from TDM could be a good source of antioxidant peptides. In addition, the antioxidant activity of hydrolyzed protein relay on protease type and hydrolysis condition.
 

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This paper introduces a technique for controlling a class of uncertain chaotic systems using an adaptive fuzzy Proportional-Integrator-Derivative (PID) controller with H∞ tracking performance. The purpose of this work is to achieve optimal tracking performance of the controller using Backtracking Search Algorithm (BSA). BSA, which is a novel heuristic algorithm, has an easy structure with single control parameter. In BSA, three basic genetic operators (selection, mutation and crossover) are utilized to generate trial individuals. To this reason, the control problem in hand is considered as an optimization problem by defining an appropriate objective function. Stability analysis of the control scheme is provided based on Lyapunov theory and modified Riccati-like equation, where the robustness of the closed-loop system is guaranteed by H∞ tracking performance for any predefined level. To evaluate the performance of the proposed control method, it is employed for tracking control of Duffing uncertain chaotic system. Simulation results show the capability of the proposed controller.  

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Hyaluronic acid (HA) is a natural and linear polymer that finds a wide-range of applications in medicine, cosmetics, and nutraceuticals because of excellent viscoelasticity, high moisture retention capacity, high biocompatibility and non-toxicity. HA has been recently produced in industrial scale by Streptococcal species. Streptococci are nutritionally fastidious lactic acid bacteria and cannot synthesize some amino acids. Therefore, it is necessary to study and select some commercial culture media for their growth. In this study, HA production and hyaluronidase activity of S. zooepidemicus ATCC 43079 in three culture media were investigated. Regarding the detrimental effect of this enzyme on HA amount, 6-O-Palmitoyl-L-ascorbic acid as hyaluronidase inhibitor was added to culture medium during fermentation. The effect of three variables consisted of glucose concentration, yeast extract concentration and medium pH each at 3 levels were considered and (response surface methodology (RSM) was used for statistical design of experiments to study the HA production by this strain. The results showed that maximum HA production was obtained when glucose concentration, yeast extract concentration and pH were 21.2 g L^-1, 43.6 g L^-1 and 6.6, respectively. Under optimum conditions, HA was produced as 370±15 mg L^-1 which was ~150% more than of HA concentration in basal medium (150±10 mg L^-1) and productivity reached 56.74 g L^-1 h^-1 that was increased 2 fold compared to central point.

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Aims: The aim of present study was production of Fatir bread fortified with rainbow trout and silver carp meats and evaluation of its quality attributes during room temperature storage.
Materials & Methods: Different concentrations of cooked rainbow trout and silver carp meat (5, 10, 15, 20 and 25%) were added to the Fatir bread and sensory properties of prepared bread were measured. Then, selected treatments were stored for 9 days at room temperature and during this time the quality attributes were evaluated. 
Findings: Results of initial sensory evaluations were showed that the Fatir bread containing 5% of rainbow trout and 10% of silver carp were accepted. Results also demonstrated that the protein, lipid and moisture content of the bread were increased with addition of the fish meat. Fatir bread fortified with fish meat had higher TVB-N, peroxide and total viable bacteria during room storage period. In terms of flavor index, the control Fatir bread was acceptable until the end of the storage period, however, the breads fortified with fish meat were within the acceptable range until day 5.
Conclusion: Can be concluded that although fortified breads showed higher nutritional value, they had lower shelf life than control bread. Between fortified breads, the quality changes were lower in the bread containing rainbow trout meat.    
 

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In order to determine the effects of fish size and weight on its salinity tolerance, the chloride cells (CCs) immunolocalization changes were examined in Caspian salmon (Salmo trutta caspius) parrs, with the same age (about 2 years old) but different types (type A: 4.88 g, 8.36 cm; type B: 14 g, 11.84 cm; type C: 24.05 g, 14.08 cm). Fish survival rate, blood osmolality, gills CCs histological and immunohistochemical changes were investigated following their transfer from freshwater (FW) to the Caspian Sea water (CSW). The survival rate increased in larger sizes and blood osmolality showed a tendency to increase in parallel with salinity. After 10 days in CSW, some abnormalities were observed in gill structure such as: lamellae cohesion, lamellae rupture and separated lamella from filament epithelium that were shown in all types. These abnormalities in type B were less than the other types. Gill CCs were observed on the gill filament and lamellae. In direct transfer of Salmon parrs to the CSW, changes in number, sectional area, and the surface occupied by CCs in the gill tissue were observed in all the three types. In the type B, the number of CCs did not change, however, in the type C, they decreased, while in the type A, there was significant increase. But in the CSW, the occupied surface by gill CCs in the type C, was reduced significantly compared to other types. According to the present results, among the Salmon parrs with the same age, the fish with type A lacked osmoregulation, while the type B had better compatibility with the CSW. However, after reaching the size of the type C and considering osmultification, it is probable that the type B fish would become compatible with the fresh water environment and they will not have osmoregulation ability in saline water for the long term.