Domestication of an endangered fish species Nandus nandus (Ham.). Pt. 1. Laboratory rearing of young fish up to sexual maturity

An experiment was conducted for rearing of Meni, Nandus nandus in laboratory condition for seven months with the objective to select appropriate feed for the species and to develop a rearing technique of the species up to the stage of sexual maturation. Different trials were conducted using artificial feed (35.5% protein), dead fresh kachki (Carica soborna), dead fresh prawn (Macrobrachium lamarrei) and live prawn (Macrobrachium lamarrei). The provision of bottom sediment did not significantly influence the growth of fish. Between dead fresh kachki and dead fresh prawn, the fish preferred dead fresh prawn. The fish was found to be reluctant to take dead fresh kachki and prawn as food unless they became very hungry. The fish was found actively feeding on live prawn. The FCR of the prawn as food for N. nandus was found to be 2.5. From the study, it was observed that in laboratory rearing N. nandus preferred live prawn as food than artificial feed, dead fresh kachki and dead fresh prawn. The fish fed on live prawn became sexually matured (eggs or white milt extruded by gentle pressure on the abdomen of the fish) in the laboratory at the end of the experiment.

Observation on the urinogenital papilla and sexual dimorphism in some Indian gobiids (Gobiidae: Teleostei)

A thorough comparative study on the urinogenital papilla and sexual dimorphism has been made for the first time in both the sexes of twelve Indian gobiids: Glassogobius giuris (Hamilton); Acentrogobius cyanomos (Bleeker); Eleotriodes muralis (Valenciennes); Parapocryptes serperaster (Richardson); Apocryptes bato (Hamilton); Scartclaos viridis (Hamilton); Boleophthalmus boddarti (Pallas), Periophthalmus schlosseri (Pallas); P. koelreuteri (Pallas); Taenioides anguillaris (Linnaeus); T. buchanani (Day); Odontamblyopus rubicundus (Hamilton). The urinogenital papilla, originating as a free muscular organ from the ventral surface of the body-wall and shortly behind anus, is present in both the sexes. It is an important organ of primary sex recognition in all species. In case of male the papilla is conical, broad at the base and in female it is either flattened, distally truncated or bluntly rounded. The presence of permanent colour mark over the specific region of the body surface is another secondary sexual character in a few species. Besides, colouration may also be a nuptial secondary sex character developed in some during peak breeding season. The enlargement and colouration of the organ is subject to seasonal variations parallel to the seasonal gonadal cycle. The histological architecture of the papilla shows a high degree of cellular specialization and an interrelationship to the urinary and genital ducts. The functional efficacy and significance of the papilla in the breeding biology of these fishes has been discussed.

Escala macroscópica de maturaçāo e ciclo sexual das espécies Penaeus indicus H. Milne Edward e Metapenaeus monoceros (Fabricius)

The macroscopic scale used for determination of sexual maturity in shrimps was tested against objective parameters namely the gonad-somatic index and the size of oocytes. The two main species caught in Mozambique, Penaeus indicus and Metapenaeus monoceros, were the object of the work. In order to reduce the subjectivity in the macroscopic exam, a table with the colours representative of each stage is presented. Since this study was conducted over a period of 13 consecutive months and there were observations from previous years, the sexual cycle of the two species is also presented, as well as an estimate of the size at first maturity.

Histology and hormonal effect of Ghrelin on ovary of Benni (Barbus sharpeyi) in order to study sexual maturity, zygote and larval generation

Benni (Barbus sharpeyi) is valuable fish that Khuzastan fisheries office propagated it artificially in Susangerd Fish Propagation Center every year. Pituitary gland is used for this aim but female fish lost their fertilization power after 2-3 years, so in present research, new hormone, that is called Ghrelin. The aims of this research are histology, hormonal, zygote and larval generation studies and comparing the results with each other. Ghrelin is a multifunctional peptidyl hormone which increases GTH-II in fish, amphibian, and birds and mammalian so its effect on Benni sexual maturation was studied. Human Ghrelin (hGRL) was obtained from ANASPEC, Canada, with 28 amino acids. In the present study, three levels of ghrelin including 0 (sham treatments), 0.10 (treatment 1) and 0.15 μg/g (treatment 2) body wt and one level of pituitary gland 4000 μg/g (pituitary treatment) with two replications were used. 56 specimens were injected intraperitonealy and their ghrelin level was evaluated immediately after injection and after 24 h. Control fish(n=16) were just injected by physiological saline. For hormonal studies sham and experimental fish(n=40) were anesthetized with MS-222 at a concentration of 250 mg l-1, and blood samples were collected and kept at 4ْC, then spun to collect serum. Serum samples were stores at -20ْC until the RIA for CTH-II. For histology studies immediately after injection a piece of ovary was collected from control fish (Sham zero) after being anesthetized. The sampled ovaries were fixed in Buin solution and embedded in paraffin, and stained to Sections of 5–6 μm using haematoxylin and eosin. The ovarian samples were performed with a compound microscope. Histology and micrometry studies had done. The mature oocytes had given from mature fish, then weighted and the working fecundity were counted. The mature oocytes fertilized, the eggs were incubated and the percentage of fertilization was calculated. After 72h the eggs hatched and the percentage of hatch was counted. The percentage of hindrance was calculated after 6 days. Hormonal results indicate that ghrelin and pituitary increase significantly the GTH-II level in comparison to sham. Macroscopic observations (before taking ovary) showed that ovaries with green colored have couple oval structure located in the abdominal cavity. Microscopic studies of dissected ovaries indicated simultaneous growth of 127 oocytes with 6 stages. The type of the ovary is asynchronous. The results indicated that both of the ghrelin treatment increased the percentage of mature follicles followed by decrease of immature follicles. There were significant differences (P<0.05) between the number of mature and immature follicles. Average diameter of follicle in both of the ghrelin treatment was significantly (P<0.05) declined in the stages of the vitellogenesis when the result compared to the other treatment. Just treatment 1 and pituitary treatment can give mature oocytes. The fecundity of pituitary treatment significantly increase in comparision to ghrelin treatment (P<0.05). In food-restricted fish where endogenous ghrelin levels are known to be increased, a chronic administration of ghrelin induces overt negative effect in releasing mature oocytes. The percentage of fertilization was significantly increase (P<0.05) in ghrelin t. in comparison to pituitary t. and the percentage of hatch was significantly increase (P<0.05) in pituitary t. in comparison to ghrelin t. There was no significant difference (P>0.05) in terms of percentage of hindrance between treatments. In conclusion, the present study demonstrated that ghrelin has positive effect on the level of GTH-II, oocyte maturation, ovarian vitellogenesis and the number of mature follicles of Barbus sharpeyi ovary. Increasing of the mature follicles number reduces their average diameter, indicating stimulating effect of ghrelin in sexual maturation of Barbus sharpeyi.The ghrelin and pituitary treatment have equal chance in the post-stage of spawning.

Study on biological characteristics, sexual maturation and reproduction of Liza abu in the water of Khozestan Province

As the most of the fish resources are known and exploited, protecting their generation is of the greatest importance. Aquaculture is one of the efficient procedures in protecting and reviving fish resources and knowing about the reproductive cycle and gonads development has an important role in approaching this aim. Liza abu belongs to the family Mugilidae that according to its resistance to the environmental condition and its fast growth , can be introduced as a fish with economical value. As there is no scientific data on the reproductive biology of this species , study on the reproductive biology and gonad development is considered as the aim of this research . For this purpose , 360 samples of this species were investigated during the period from February 2007 to January 2008 in Khozestan Province . After studing morphological and histological characteristics of gonad specimen , they were prepared through histological method. Samples were prepared through usual histological method and studied under light microscope. According to the results, the maturity stages of male and female Liza abu were separated to six different successive stages. In ovaries , these stages were as follow : In stage І, the oocytes were small , this stage was observed from July to October . In stage ІІ, considerable growth was observed in the oocytes . This stage was observed from October to January . In stage III, due to vitellogenesis, the maximum growth was observed and three layers of theca, granullosa and follicle cells were visible. This stage was observed during January and February . In stage IV, migration of germinal vesicle was observed and due to hydration of the oocytes , their diameter was increased. The ovaries were yellowish and in maximum size and ovules could be easily observed with naked-eye . This stage was observed in February and March . In stage V, spawning occured. This stage was observed in April . In stage VI, ovaries consisted of immature and atretic oocytes and also empty follicles. This stage was observed in May and June. In testes , these stages were as follow : In stage I , the testes were small in size and contained the spermatogonia which were the only cellular components.This stage was observed in August and September . In stage II (maturing virgin ) , the spermatogonia and the primary spermatocytes were visible. This stage was observed in October . In stage III (developing), intensive spermatogenesis was occured and the primary and the secondary spermatocytes were the most visible cells during this stage .This stage was observed from November to January. In stage IV(developed), cells of all stages of spermatogenesis could be seen but the secondary spermatocytes and spermatids were in large number. This stage was observed from January to March. In stage V , the testes were filled with sperms. This stage was observed in March and April .In stage VI, residual spermatozoa and the spermatogonia were visible in the testes. This stage was observed from May to August. According to cyclic changes in GSI, sexual maturation in breeding begins in January and spawning occurs in April. The ova diameter ranged from 30.75 μ in stage I to 472.19 μ in stage IV. In this study , the sex ratio was 1:2.7, and male and female percentage were 27.02% and 72.98% respectively. This means that females predominate males. In this study absolute fecundity was calculated and changing between 30805.44 to 431247.3 was observed and absolute fecundity was calculated 111275.3 in average.

The survey of sexual maturity in Mugil cephalus so measuring sex hormones and histologic studies

In this study the process of female gray mullet brooders was carried out by using histological study and masurment of sex steroids. Results of histological studies showed that oocyte of gray mullet brooders in Gomishan Rearing Center conditions of develop to the end of yolk globule stage. The results were observed with oocyte in chromatin nucleolar stage (first stage) with means of diameter of 20 p m, in August, perinucleolar stage (second stage) in September with mean diameter of 87 p m, yolk vesicle stage (third stage) in October with mean diameter 200 p m and yolk granules stage (forth stage) from October to November with average diameter of 180 — 650 p m. For the reason of stopping oocyte develop at the end of fourth stage, hormonal induction to final oocyte maturation and ovulation was used. For this purpose, carp pituitary , HCG and LRH-A2 with different combinations were used in two stages, second injection was used 24 hours after first injection. 15 females brooders were divided in 5 groups, different hormonal combinations were injected to four groups and to fifth group as control, only saline, was injected. The process of female brooder rippening in hormonal induction was studied via masurment of sex steroids including 17 a - hydroxy progestrone, estradio1-17)6 and testosterone. Blood samples were collected from caudal vein during first injection, 24, 30 and 48 hours after the first injection. At the same time, for distinguishing histological changes the sample has been attained from the gonads Sex stroid fluctuation patterns in different brooder groups that injected hormon were similar, however hormonal composition had similar effects. All brooder that their oocyte in the beginning of hormonal injection were At the end of fourth stage with oocyte diameter average of 600 p m received to final maturation and ovulation. The brooder that its oocytes were At the begining or mid-fourth stage did not show ovulation but hormonal induction caused oocyte develop at the beginning of fifth stage. Study of 17-hydroxy progestrone fluctuation showed that the maximum level of this steroid (0.347 ng/ml) measured 30 hours after the first injection and was significantly higher (p< 0.05) than those of control group. So, 17-hydroxy progestrone is probably precursor of maturation inducing steroid (MIS). However the maximum level of that observed was coincident with germinal vesicle breakdown, oil droplets coalescence and dissolution of yolk granuls The maximum levels of esteradiol— 17/0 and testosterone (3.778 and 16.801ng/ml,respectively) in spawned brooders,were observed 24 hours after the first injection. levels of those steroids were significantly higher (p<0.05) than control group. Maximum level of sex steroids in the brooders that did not spawn to the end of treatment was observed with more delay than those in spawned brooders. Therefor maximum level of 17a-hydroxy progestrone (0.264 ng/ml) in those brooders observed in fourth sampling time and the maximum levels of estradio1-17a and testosterone (2.944 and 18.993 ng/ml, respectivly)observed in third sampling time that was significantly higher (p<0.05) than those of control group. For the study of stress effect on brooders during the hormonal induction, level of cortisol was measured in every sampling time. level of cortisol had high fluctuation that showed handling level and stress effect on brooders. However maximum level of cortisol in majority of brooders was dominant in third sampling time that was coincident with final maturation.

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.