Reproductive biology, artificial propagation and larval rearing of two freshwater eels, Monopterus cuchia and Mastacembelus armatus

Studies on reproductive biology and artificial propagation including larval rearing of freshwater mud eel, Monopterus cuchia and spiny eel, Mastacembelus armatus were attempted. The gonadosomatic index (GSI) of mud eel ranged from 0.41 (August) to 5.52 (June) in males and 0.53 (August) to 7.61 (June) in females. In both cases the GSI showed a peak in June. Fecundity ranged from 228 (TL - 396 mm; W - 78g) to 5510 (TL - 865 mm; W - 630 g). In case of spiny eel, the GSI varied from 0.65 (August) to 8.30 (July) in males and 0.70 (August) to 10.46 (July) in females. GSI showed single peak in July. Fecundity ranged from 570 (TL - 240 mm; W - 30 g) to 10870 (TL - 601; W - 350g). Histology of the testes and ovaries of the eels were carried out to investigate the gonadal development stages during the reproductive months (August to November 2003). In case of male M. cuchia, the secondary primordial germ cells, primary spermatogonium, some spermatogonia A and clone of spermatogonium B in testis were observed in September. In October-males different sized lobules having spermatogonia, spermatocytes and spermatids were observed. In the ovary of M. cuchia, polygonal shaped oocytes were seen during September. The oogonia were reduced with dense and irregular shaped during October. Numerous pycnotic cells were visible during November. In male M. armatus numerous broken lobule walls were found in testes during September. In October, abundant primary germ cells, pycnotic nests of degenerating cells, spermatogonia and spermatids were observed. In females, ovaries had distinct yolk vesicles stage and yolk granules stages in August. In September, the follicular cells of the oogonia were ruptured, shrunk forming irregular shaped in October. Oogonia were also shrunk with thin, irregular shaped structure but broken parts of the ruptured follicular cells were scattered in case of M. armatus. Experimental attempts on artificial propagation indicated that both freshwater eels were difficult to breed using inducing agents like pituitary glands (PG) of 10, 20, 50, 100 and 150 mg per kg of body weight. Same doses were used for both sexes with equal sex-ratio. In both cases, brood fish died at higher doses of injection given at 100 and 150 mg PG/kg bodyweight. However, M. cuchia breed naturally in cisterns when provided with water hyacinths and tunnel in muddy bottom. M. cuchia fed with chopped cooked fish attained a mean weight of 18.75 ± 2.3 g and cent percent survival. While in case of M. armatus best growth by weight (12.0 ± 2.48 g) and cent percent survival were achieved using chopped raw fish. Car tyre was observed as best shelter for attaining the mean weight gain 22.53 ± 2.24 g and cent percent survival of M. cuchia. While PVC pipe was found to be the best shelter for M. armatus, where it attained the mean weight of 12.73 ± 1.88 g and cent percent survival.

Larval development of a semi terrestrial mangrove sesarmine crab Chasmagnathus convexus (Crustacea: Decapoda: Brachyura: Grapsidae) reared in laboratory

Four zoeal stages and one megalopal stage were identified in laboratory reared semiterrestrial mangrove sesarmine crab Chasmagnathus convexus. At an average salinity and temperature of 20±1% and 19.2±0.2°C, the megalopa was attained 24 days after hatching. Morphologically, the first zoae of C. convexz1s is very similar to those of other species of the genus Chasmagnathus as well as species of the genus Helice, in that view all share the following characteristics: lateral spine on the carapace, three pairs of setae on the posterior margin of the telson furca, one plus five setae on the endopod of the maxillule, and two plus two setae on the endopod of the maxilla. The differences between the first zoea and megalopa of and those of its congeners are discussed.

Larval rearing and fry raising of Pangasius sutchi(Fowler)

Pangasius sutchi were artificially bred for determining the hatching success and larval growth response to live food in relation to varying stocking densities. The fertilized eggs were hatched out with successful hatching rates ranging between 60 and 63%. Newly hatched larvae of 4.4 mm average length were reared using Tubifex as live food in metallic trays with water temperature of 27 to 29.5°C and dissolved oxygen level of 3.88 to 6.22 mg/1 for 6-day with an average survival rate of75.56±13.25%. The P. sutchifry of9- day old were further reared using Tubifex in the polythene covered metallic trays at the stocking densities of 2-7 fry per litre of water for a period of 14 day. P. sutchi fry raising at 4 individual per litre of water for 14 day gives better results in terms of survival and growth.

Observations on the ingression and distribution of larval prawns in the Nethravati-Gurupur estuary, Mangalore

The spatial and temporal distributions of larval prawns of penaeids and non-penaeids in the estuarine waters of Mangalore were studied. Larvae appear to be passively brought in by the incoming flood tides to the estuary, enjoy a wider distribution throughout the estuarine complex with abundance towards the mouth. The distribution of larval prawns was more in Nethravati than in the Gurupur stretch. The influence of temperature, hydrogen-ion-concentration, salinity and dissolved oxygen on the distribution of larvae in the estuaries is discussed. Inference on spawning seasons of commercially important prawns in the neighbouring waters has been arrived at based on their larval abundance.

Experiments on the induced spawning and larval rearing of the milkfish, Chanos chanos (Forsskal) in 1979

This paper summarizes the results of the experiments on the induced breeding and larval rearing of milkfish (Chanos chanos) during the 1979 season. Milkfish larvae could be reared successfully without the use of trochophore larvae of oysters as feed during the first few days. In order to induce the ovulation of wild adult milkfish a higher dose of human chorionic gonadotropin hormone is required.

Effect of furanace on the development of larval stages of Penaeus monodon Fabricius

Zoea 2(Z SUB-2 ) Mysis 1 (M SUB-1 ) and Postlarva 1 (P SUB-1 ) of P. monodon artificially spawned in closed-system concrete hatchery tanks were bioassayed for their tolerance to the antibiotic furanace. The setup consisted of four 20-liter capacity plastic basins previously conditioned for 15 days with freshwater in full sunlight. During the experiment, each basin was filled with 5 liters of seawater to which was added filtered Chaetoceros and Brachionus to give densities of 5 . 0-7 . 5 x 10 SUP-4 cells/ml and 10-20 individuals/ml, respectively. The following are the properties of the water used throughout the experiments: salinity, 26-32%; pH, 7 . 3-8 . 4; temperature, 25-30 degree C; dissolved oxygen, 4 . 5-8 . 4 ppm; nitrite, 0 . 36-0 . 99 ppm; and ammonia, 0 . 10-0 . 30 ppm. To each basin were added 50 healthy larvae of specific stages of P. monodon. After an initial acclimation of one hour in the medium, preweighed amounts of the antibiotic were added and thoroughly dissolved. The concentrations tested were 1 . 0, 2 . 0 and 3 . 0 ppm. One basin always served as control. After 24 hours of exposure, the surviving population in each basin was counted. The survivors were then examined thoroughly under the microscope for unusual behavior and morphological defects brought about by the exposure. To minimize wide variations in the medium as a result of feeding and other manipulations, the systems were all prepared at 9:00 a.m. each time, and the feeds on two instances, one at 5:00 p.m. and another at 5:00 a.m. Fifteen trials conducted with Z SUB-2 showed survival ranges of 68% to 98% with a mean of 77 . 6% in the controls; 32% to 94% with a mean of 65 . 7% at 1 ppm, and 0% to 56% with a mean of 36 . 5% at 2 ppm. There were no survivors at 3 ppm. Interpolation from the survival-dose curve gave a 24-hr LC SUB-50 of approximately 1 . 6 ppm.

Observations on the artificial fertilization of eggs and larval rearing of the grey mullet, Mugil cephalus L.

The results are presented of attempts to artificially fertilize Mugil cephalus eggs in the Philippines. Embryonic development is outlined and rearing of the larvae described. Mass mortality occurred during week 3 of rearing.

Notes on the pigmentation pattern in the larval developmental stages of laboratory-reared milkfish

Milkfish fry were artificially bred and reared in the laboratory and the pigmentation pattern of the different developmental stages of the larvae are described in detail, with illustrations.

Experiment on larval rearing of Penaeus monodon fed with cultured diatom Chaetoceros calcitran and fermented vegetable trash

A brief account is given of experiments undertaken rearing Penaeus monodon larvae fed on diatom (Chaetoceros calcitran) and fermented vegetable trash, which included fruits and their peels, vegetables and rice. The possible use of high protein content trash materials as a feed substitute is examined briefly.

Viability of frozen algae used as food for larval penaeids

Freezing with added chemicals as flocculants and protectants was assessed as a means of preserving stock cultures of 4 algal species used for larval penaeid food Chaetoceros calcitrans, Skeletonema costatum, Tetraselmis chuii and Isochrysis galbana . The maximum storage effectability of the preservation techniques for each species was also determined.

Growth phases of cultured algae used as larval food

A study was undertaken to determine the onset and duration of the growth phases of cultured algae commonly used as larval food (Skeletonema costatum, Chaetoceros calcitrans, Tetraselmis chuii, Chlorella vulgaris, Isochrysis galbana) so as to predict the time of harvest at the desired stage to suit various needs and purposes.

Bacteria from seawater used in Penaeus monodon larval cultures

Bacteria in the seawater used in P. monodon hatchery operations were isolated on Bachmann's agar. The total plate counts in 25 isolations ranged from 1.0 - 5.0 x 10² to 5.1 -10.0 x 10⁵ cells per ml. Out of 124 isolates, 98 (79 percent) were Gram-positive and 26 (21 percent) were Gram-negative. Micrococcus and Staphylococcus were dominant in the former group, while Acinetobacter, Moraxella, Flavobacterium and Alcaligenes were most numerous in the latter. Twenty-nine of the Gram-positive isolates closely resembled Peptostreptococcus, Planococcus, and Pediococcus.

Viability of frozen algae used as food for larval penaeids

Freezing with added chemicals as flocculants and protectants was assessed as a means of preserving stock cultures of 4 algal species used for larval penaeid food Chaetoceros calcitrans, Skeletonema costatum, Tetraselmis chuii and Isochrysis galbana. The maximum storage effectability of the preservation techniques for each species was also determined.

Embryonic and larval development of Mystus gulio (Ham.)

Mystus gulio eggs are strongly adhesive and contain relatively small yolk (0.75-1.0 mm). The egg envelop is thick and transparent. First cleavage (two cells), four cells, eight cells, sixteen cells and multi cells stages were found 20, 25, 35-40, 60 and 70 minutes after fertilization, respectively. The morula stage was visualized within 1.5 h after fertilization. The heart beat visible and the circulatory system commenced after 16 h of fertilization. Embryos hatched 18-20h after activation of egg. The newly hatched larva measured 2.82±0.03 mm in length and 0.32±0.06 mg in weight. The yolk sac was fully absorbed by the third day though larvae commenced exogenous feeding even before completion of yolk absorption. A 5-day old post larva began wandering in search of food. Ten-day old post larvae endowed with eight branched rays in dorsal fin and seven in caudal fin. Fifteen-day old post larvae had the pectm:al spine become stout though the embryonic fin folds had to be disappeared. The length of fingerlings ranged from 25-30 mm after 30 days, and their external features were just like those of an adult except that they were not sexually matured.

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.