Purification and properties of Aspartate aminotransferase (E.C. 2.6.1.1) from skeletal muscle of Cirrhina mrigala

Aspartate aminotransferase (E.C. 2.6.1.1.) from the skeletal muscle of fresh water fish Cirrhina mrigala has been purified 40 fold by ammonium sulphate fractionation, adsorption on alumina Csub(8) gel and chromatography using DEAE-cellulose column and the properties of the purified enzyme studied. The pH optimum of the enzyme is 7.8. The Km value of aspartic acid and 2-oxoglutaric acid are found to be 2.8 x 10sub(-3) M and 1.0 x 10sub(-4) M respectively. The activity of enzyme is inhibited by p-chloromercurybenzoate, hydroxylamine hydrochloride and sodium cyanide. The inhibition by pchloromercurybenzoate is reversed by reduced glutathione, B-mercaptoethanol and cysteine. Dicarboxylic acids such as maleic acid, malic acid and succinic acid inhibit the enzyme activity. The enzyme is not activated by any of the metal ions tested and heavy metal ions such as mercury and silver strongly inhibit the enzyme activity.

Survey of biosystematic and sexual reproductive physiology in Holothuroidea on Hormozgan Province

Sea cucumbers belong to phylum Echinodermata, order Holothuroidea are an abundant and diverse group of Invertebrates, with over 1400 species occuring from the intertidal to the deepest oceanic trenches. Sea cucumbers are important components of the food chain in temperate and coral reef ecosystems and they play an important role as deposite feeders and suspension feeders. Rapid decline in populations may have serious consequences for the survival of other species that are part of the same complex food web,as the eggs, larve and juveniles constitute an important food source for the other marine species including crustaceans, fish and mollusks. In addition sea cucumbers are often called the earthworms of the sea, because they are responsible for the extensive shifting and mixing of the substrate, and recycling of detrital matter. Sea cucumbers consume and grind sediment and organic material into finer particles , turning over the top layers of sediment in lagoons , reefs and other habitats and allowing the penetration of oxygen. While the taxonomy of the holothurian families is generally well known , the distinction of similar species is difficult. There are relatively few holothurian taxonomist.Most sea cucumber species can be identified by Holothurin taxonomists by using the calcareous skeletal ossicles found in the body wall. In this study , at first a sea cucumber from Kish island in Persian gulf has recognized. Individuals collected from west and east extend far away into north and south of coral reefs by diving. I have checked them morphologically and anatomically.Then with key to the orders of the Holothuroidea, They belong to the Aspidochirotida with key to the families of Aspidochirotida, they were in Stichopodidae families and with key to the genus of Stchopodidae, they were Stichopus. Then ossicles were extracted at National Museum of Natural History, by Dr David Pawson. The ossicles were measured on a transect across a slide prepared from the mid-dorsal region of each specimen.The one we have in the shallow waters of Kish island, is Stichopus hermanni, a massive holothurian, body broad, considerably flattened ventraly ,the dorsal side slightly arched and the lateral sides almost vertical; body wall fairy thick and soft ; mouth subterminal; anus central; tentacles usually 20 in number of length and leaf shaped. Numerous ossicles consisting of table with large discs having usually 7 to 15 peripheral holes, but often irregular or incomplete and spire of moderate height ending in a group of spinelets, rosettes of variable development, and c-shaped rods. Color (exept papillae)partly remained after preservation in alcohol which is found at the depth of 4 to 8 meters, on coral reef. Furthermore, the sexual reproductive cycle was described using standard methods. Gonads were removed and transferred to Bouin's fixative for four weeks and then processed according to standard embedding technique. To prevent the loss of tubule contents during embedding, the tubule sections, were cut well beyond the segment selected for sectioning. For each individual, six sections, each section with 5µm diameter by microtome were cut from tubules. These sections were first placed on gelatin coated slides (the gelatin was heated to 42°c) and then transferred to the oven at 37°c for one hour. This technique usually prevents the fragil tubules from breaking and the loss of gametes. The slides were stained with Eosin and Hematoxylin, and good resolution of the various cell types achieved.A second series of slides was stained with the Periodic Acid Schiff(PAS) to identify polysaccharides(glycogen). Monthly sampling was occurred.The sexual reproductive cycle was defined through the combined use of these criteria: Monthly percentages of the gonad stages for each sex, the monthly gonad index (GI) , given as the ratio of the wet gonad weight (G) to the dray weight (DW)and the monthly percentage of individuals that undetermined sex. The gonad consists of two tufts of tubules on which saccules develop. Gonadal development was classified into five stages: post spawning, recovery, growth, advanced growth, and mature stage that were adapted from the earlier studies of holothurians. Histological preparations showed that the sex of larger individuals could be identified by the presence of oogonia and young oocytes in females, and spermatogonic stages in males.The mean diameter of the tubules and gonadal mass follow annual cycles, increasing from late winter through spring, and dropping abruptly after spawning in the summer. Gametogenesis is generally a prolongate process and begins in March. By summer the ovarian tubules contain oocytes with diameter of 120-240 pm and the testicular tubules contain an abundance of spermatozoa (diameter 5-6 gm ).Following spawning the predominant activity within the spent tubules is phagocytosis of the residual gamets.The active phase of gametogenesis (March to July), coincides with an increasing photoperiod regim, and an accelerated gametogenesis occurs in July when temperature is high. Throughout the year, the gonad of Stichopus hermanni is larger in males than in females, and this is due to the number of tubules in the testis rather than to tubules length or diameter.

Reproductive physiology of ribbon fish (Trichiurus lepturus)

The ribbon fishes ‘of the family Trichiuridac are represented as one of the most important food resources in Indian ocean. High density of the dominant species of ribbon fish (Trichiurus lepturus) in Oman sea and the 'Tillable catch in last yeas (more than 7000 tones per year) makes a trust area for studing their population biolog and stock assessment. As our knowledge on reproductive biology of this species has an important role on their fisheries management, as well as conservation of this stock from decline or over fishing, this research was held to determine some aspects of reproductive physiology of ribbon fish and the effects of environmental factors in gonadal cycle. The goals of the present thesis is to determine some aspects of reproductive physiology such as gonadosomatic index (GSI) , hepatosomatic index (HSI), condition factor (Ko, fecundity, sex ratio, size at first maturity, size at maturity (LM5O) and their relative hormonal & biochemical fluctuations. In this regards annual variation of sex hormones ic. estradiol 17-B, progestron, cortisol, testostrone and gonadotropins FSH (GTH-I) , LH (GTH-ll)I were measured ; gonadal histological studies were done by light & electron micrography. The research was carried out from April 1995 to January 19% in Ras Nleidani in the north part of Oman sea, and the environmental factors such as temperature, salinity, oxygen, rainfall and pH were measured. The effects of these parameters on reproductive cycle and hormonal fluctuationswere discussed by using correlation and principle component analysis (PCA). Female Ribbon fish reproductive strategy shows the same paterns of nonguarder marine teleosts. T. lepturus has more than one spawning season (existance of egges in different size in each month) and therfore it must have asynchronous ovaries and belong to continious spawners. GSI and HSI are good evidences for this type of reproductive patern. The testis of the lobular type , which is typical of most teleosts , is composed of numerous lobules which are separated from each other by a thin layer of fibrous connective tissue. GSI fluctuations revealed prolong- spawning time in males. There is significant increase in 17-13 estradiol. progestrone , cortisol and gonadotropins with maturity and prespawning period of female T lepturus. Plasma concentration of E2 and GTH II incresaed along with water temperature increasing (3300).. Spawning was observed from Nov. 1995 to Apr. 1996 in this species. Progestrone increased significantly with increasing rainfall in this season (P<0.01). Plasma cortisol levels increased with maturation and vitelpgenesis and also with the peak of spawning. From lenght-weight frequency and size distribution in each age groups and also minimum size at first maturity (52a cm) it would he concluded that T. lepturus must be matured at 2 years of age. Serum cholestrol and triglicerides significantly increased when maturation occured in this species. The relationship between alkaline phosphatase activity and hormonal fluctuations with maturity and vitelogenesis were discussed. Proximate compostion (muscle) shows significant variation with spawning period and maturity. Absolute individual fecundity (17420-159150) increased with body length and weight. Ultrastructural observations show dramatic variation in cell membrane (0ocyte membrane), yolk vesicles and, nucleolus dispersal in relation to maturity stages. fluctuations of gonadal hormones were discused in relation with vitelogenesis. Testosterone increased in males from Nov: to Mar. due to environmental impacts and spawning time. Sex ratio in different depth (10-40 m ,80-110 m) shows significnt differences in this ratio for two depths. In 10-40 m depth female shows dominant abundance to male in each months that may be due to their reproductive migration behaviour. The effects of temperature photoperiod and rainfall to maturity and spawning were discussed. According to -pawning period of T. leptunts in our sampling area it could be suggested that ribbon fish fi,theries must be restricted in the peak of spawning seasons (Feb. to Mar.) and in the spawning grounds (under 40 m depths). Other suggestions for population conservation have been mentioned.