Development of artificial breeding techniques for long-whiskered catfish, Sperata aor and giant river catfish, Sperata seenghala of Bangladesh

Sperata aor and S. seenghala are the two important native catfishes of Bangladesh but commercial farming of these species is not possible due to lack of naturally collected or artificially produced seeds for stocking. Attempts were made to develop techniques for seed production by artificial breeding and nursery-rearing of fries of these catfishes. A total of 60 S. seenghala (750-1,500 g) and 10 S. aor (600-1,000 g) broods were collected from the Brahmaputra river-basin and floodplains in Mymensingh region four months prior to their breeding season. The collected brood fishes were reared in separate earthen ponds with supplementary feeds comprising of rice bran (40%), mustard oil cake (29%), fish meal (30%) and vitamin-premix (1 %). Three experiments were conducted to optimize the hormone dose. A total of nine S. seenghala females weighing from 750 to 1,500 g were given an initial and resolving dose of 12-20 and 16-24 mg PG/kg body weight, respectively. The males weighing from 650-950 g were administered a single dose of 18-26 mg PG/kg body weight at the time of the time of administering the resolving dose to the females. The females ovulated partially and the eggs were examined under a compound microscope, but most of them were found to be less ripe or damaged. Collection of milt by stripping the males was not successful. The testes were taken out and sperm were observed to be non-motile and less developed. In view of stimulating natural propagation of S. seenghala, artificial holes (nests) were constructed in the pond bottom. Each hole was 0.7 m in diameter and 0.3 m in depth. A total of 10 holes were made and then 10 pairs of S. seenghala breeders (800-1,200 g) were stocked in the pond. In mid February, 3,000 fry of S. seenghala with a mean length of 4.60 cm and weight of 0.36 g were collected by repeated netting followed by drying of the pond. The fry were then stocked in a nursery pond and fed with commercial feed (SABINCO starter-1). The average length and weight of the fingerlings were 9.01 cm and 3.95 g, respectively and the estimated survival was 60% after two months of rearing. S. aor did not respond to natural spawning. Further study is essential to develop techniques for their successful artificial and natural breeding.

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.

Tolerance of Penaeus monodon larvae to cupric sulfate added in bath

Copper is used to deter the growth of bacterial, fungal and protozoan disease organism in fishes. Zoeae (Z SUB-1 ), myses (M SUB-1 ) and postlarvae (P SUB-1 ) were exposed to copper sulfate at concentrations of 0 . 025, 0 . 05, 0 . 75, 0 . 1 and 0 . 2 ppm from 24 to 96 hours. The number of surviving larvae were counted at the end of each 24-hour period and the percentage of survival is determined for each dose level. The LC SUB-50 for each of the larval stages was interpolated from the data whenever possible. Three trials with 2 replicates per trial were conducted. The physico-chemical characteristics of the bath taken before and at the end of the experimental period show insignificant differences between initial and final values in each trial. Results indicate that mortality rates of all larval stages increased with exposure time and that mortality rates of the experimental group is higher than the control. Interpolation of the LC SUB-50 is possible only for the 48-h and 72-h exposure times for both zoeae and myses and for the 48-h exposure time for the postlarvae. This is due to the high survival percentage of the 24-h group and the low survival percentage (below 50%) of the larvae exposed for 96 hours. The 48-hour LC SUB-50 for Z SUB-1 , M SUB-1 and P SUB-1 are 0 . 225, 0 . 350 and 0 . 125 ppm respectively. Postlarvae seem to be more sensitive than either of the 2 larval stages having a lower 48-h LC SUB-50 and a low survival rate after 72 hours. The larvae were observed to lose their balance and were lethargic, producing few swimming movements so that they were mostly confined to the bottom of the aquaria. Moribund larvae observed under the microscope had a faster but weak heartbeat compared to healthy larvae. Slight or complete loss of feeding ability indicated by empty guts and delayed molting of Z SUB-1 to Z SUB-2 were also noted.

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.

Survey of reproductive physiology on Epinephelus coioides in Khozestan province, Persian Gulf

Epinephelus coioides (family serranidae) is protogynous. This species is one of the most important fishes in food chain of marine proteins of persian Gulf. Therefore knowing about the reproductive biology and physiology of this species is an important role on aquaculture procedures. Monthly samples of Epinephelus coioides were obtained in khozestan Bahrekan province from 2001 to 2002 for annual variation of base line of reproductive hormone. The hormones such as: 17-B estradiol, Testosteron, Progesterone, Gonadotropin I ,II GTHI, II) and cortisol have assayed and also different stages of gonads from the histological point of view were studied by light and electron microscope. Aditional to morphometric and fecundity measurements, the important factors such as : Gonadosomatic index (GSI) Hepatosomotic index (HSI) and Condition factor (KF) were also studied. Environmental factors such as temperature, salinity, photoperiod and pH were analyzed for the determination of effective factors responsible for the changes of reproductive cycles. The flactmation of estroid hormones and gonadotropines show a significant variation in different stages of maturation, e.g 17-B estradiol's concentration in the third stages, GTH II in fourth stages of sexual maturation or final oocyte maturation, plasma Testosteron in post ovulation and Progesterone during maturation indicates the highest levels of above mentioned hormones. The total calcium concentration was high in all year. calcium concentration was correlated with GTH II synthesis and increases with GTH II in June. 17-B estradiol concentration was also correlated with GSI. The high concentration of cortisol throughout the year was an index of stress and development of ovary maturational processes. This species was protogynous synchronous hermaphrodites , and belongs to annual spawning species, being monandric. The sexual transition was found to occure in individuals of 51.2- 105 cm in length. GSI and HSI level confirms the time of spawning period is in April- June. Electrone microscopic studies of gonad tissues showed some changes in mitochondria and endoplasmic reticulum in the post ovulation, maturation and post spawning periods. During the monthly sampling the biochemistry of tissues variations indicated decrease in protein and lipid content, but an increase in water content of spawning fishes which was correlated to the maturation of Epinephelus coioides . sex ratio indicative of higher frequences of females to males during monthly sampling periods. The females were smaller than males in sizes, therefore the females lived in 8-15m depth, but males were living in upper limits of depth. The results indicated that the temperature was the most effective parameter in reproductive cycle of Epinephelus coioides and the mean 24°c was a convenient temperature for spawning. Photoperiod was the second effective. factor on the reproductive cycle for this species. It seemed that the increase in the photoperiod between January to May caused a development of the oocyte. Regarding to the results of this research, it seems that the period of spawning in Epinephelus coioides is in May- June and the aquaculture procedure of Epinephelus coioides could be performed in the above mentioned periods.

Study ectoparasite infestation of 10 common imported freshwater ornamental fish species

In the present study, ectoparasite infestation of common freshwater ornamental fish species imported into Iran including: Dwarf gourami (Colisa lalia), Angelfish (Pterophyllum scalare), Oscar (Astronatus ocellatus), Red eye Tetra ( Moenkhansia Sanctaefilomenae), Barb( Capoeta tetrazona), Arowana (Osteoglossum bicirrhosum), Goldfish(Carassius auratus), Red fin Shark (Lebeo elythrurus, Catfish (Hypostomus plecostomus) and Pangasius sutchi ( Pangasius hypophthalmus) from May until April was investigated.A total of 6 specimens sample of each fish species) randomly were obtained and taken alive to the lab. After observing gill and skin wet smear under the microscope , gill was dissected and examined carefully .Photo and films were taken from the isolated parasites and parasite identification was performed according to Yamaguti AP1), Bychowsky (■Y).From a total of examined fishes 1\ ► sample (YY.V.) were parasitized .Parasite groups that were observed consisting ciliated protozoan myxosporidian , monogenean Digenean metacercaria and crustacean .The only non-parasitized fish was Barb. Monogenea Trianchoratus sp. , Cleidodiscus sp. , Ancylodiscoides sp. , Thaparocleidus sp. , Centrocestus formosanus metacercaria and Myxobolus longisporus report for the first time in ornamental fish which imported from Southeast of Asia to Iran. According to the results, it seems that severe quarantine and sanitary rules are necessary.