Studies on the egg quality of Penaeus monodon Fabricius based on morphology and hatching rates

Eggs of P. monodon are classified into 5 different types on the basis of morphological criteria and hatching rates: A sub(1) eggs, which undergo normal development with 58% hatching rate; A sub(2) eggs, which show delayed and/or abnormal development with 32% hatching rate; B eggs, which are unfertilized and characterized by irregular cytoplasmic formation; C eggs, which are unfertilized and show no change in appearance; and D eggs, which are unfertilized and show extensive bacterial presence. The establishment of a highly linear relationship between percent A sub(1) eggs and hatching rate provides a useful tool to aid the hatchery technician in determining which spawnings to reject outright and which to rear through the larval and post-larval stages.

Effect of substrate types on fecundity and nauplii production of ablated Penaeus monodon Fabricius

Experiments were undertaken to assess the survival, spawning, fecundity and nauplii production of ablated P. monodon females reared in flow-through broodstock tanks with white coralline and black sand substrate for 62 days. The similar trend observed in mortality rates in both substrates suggests that variation in substrate material for broodstock tanks is not a likely cause of prawn mortality. There were also no significant differences observed between rematurtion rates, i.e. number of spawnings, under the different treatments. Singnificantly higher nauplii production were observed in females in tanks with white substrates. At present, the land-based broodstock tanks in SEAFDEC utilize white coralline substrates due to higher hatching rate of eggs and nauplii production, convenience in siphoning out debris and excess food that tend to accumulte in the tank, and contrast provided by the white substrate during nightly observations of ovaries.

In vitro effects of fungicides on the fungus Haliphthoros philippinensis

Pure cultures of Haliphthoros philippinensis, isolated from infected Penaeus monodon larvae, were exposed for 24 hours to varying concentrations of antifungal agents. The efficiency of each agent to inhibit sporulation and mycelial growth was measured. Effects on P. monodon eggs and larvae were also investigated. It is concluded that preliminary bioassay of larval tolerance to the suggested effective doses should always be made prior to prophylaxix or therapeutic applications.

Effect of different sex ratios of ablated wild-stock Penaeus monodon Fabricius on maturation, fecundity and hatching rates

The study was conducted to determine other sex ratios of ablated wild-stock Penaeus monodon other than the most commonly practiced 1 male: 1 female. Four different sex ratios, 0:1, 1:1, 1:2, and 1:4 male: female were tested in four 4m diameter circular tanks for a period of 55 days. During the first run the 1 male: 2 females ratio gave: (a) the highest percentage of first (42.20%), second (30.00%) and third (33.33%) spawning; and (b) the highest total and average fecundity (3.9 million eggs and 300,692 eggs, respectively). The 1 male: 2 females ratio is recommended on the basis of highest percentage for the first, second and third spawners, total and average fecundity.

Survival, maturation, fecundity and hatching rates of unablated and ablated Penaeus indicus H.M. Edwards from brackishwater ponds

Penaeus indicus harvested after three months of rearing in brackishwater ponds and averaging 6.9 g for females and 5.6 g for males were stocked in two 12 cu m flowthrough ferrocement tanks at 240 females and 200 males per tank. The females were ablated on one eyestalk in one tank and remained unablated in the other tank; all males were unablated. Ablated females spawned up to 7 times per female; unablated females spawned up to only 3 times during the two month duration of the experiment. Ablated females produced a total number of 17.5 x 10⁶ eggs, 6.6 x 10⁶ nauplii, and an average of 23,480 eggs/spawning and 37.8% hatching rate from a total of 757 spawnings. Unablated females produced a total of 2.0 x 10⁶ eggs, 1.1 x 10⁶ nauplii, and an average of 26.990 eggs/spawning and 53.9% hatching rate from a total of 74 spawnings. Survival of ablated females was 53.5% compared to 69.4% for unablated females; males in both tanks averaged more than 90% survival.

Captive breeding and embryonic development of honey gourami, Colisa sota (Ham.-Buch.)

Honey Gourami, Colisa sota, has high ornamental as well as food value. The natural resources of this species are gradually declining, due to destruction of its habitat, over fishing for aquarium trade and human consumption. The fish was bred in captivity under controlled environment. It laid about 200-400 eggs in bubble nest built by the male. Hatching started within 28-30hrs. after egg laying. The hatchlings became free swimming by 3rd to 4th day of hatching. The male showed territoriality and parental care by guarding the eggs and hatchlings. The larval survival was 30-35%. The breeding behavior, embryonic and post embryonic development of the fish were studied.

Spawning behaviour and induced breeding of an estuarine catfish, Mystus gulio (Ham.)

Spawning behaviour of hormone induced estuarine catfish, Mystus gulio was observed in captive condition. Spawning activities that include pairing, chasing and resting, nudging, and twisting, started about 5 hours post injection and ended with release of eggs within 1-2 hours of courtship. Three different dosages of "ovaprim" (1 ml/kg, 1.5 ml/kg, and 2 ml/kg in a single dose) were used in induced breeding of M gulio. The latency period was less (6-7 hours) with the dose of 1.5 and 2 ml/kg, while it was more (7-8 hours) with that of 1 ml/kg. However, all females spawned successfully with each of three different dosages, without any significant differences in the rate of fertilization and hatching. Eggs under all hormone dosages hatched between 18-20 hours after spawning. The hatching rate with 1, 1.5, and 2 ml/kg varied from 71.3-72.7%, corresponding to the fertilization rate of 80.7-84.7%.

A comparative study on the induced breeding and in vitro fertilization performance by various inducing agents in catfish, Heteropneustes fossilis (Bloch)

Brood catfish, Heteropneustes fossilis were collected from Tamirabarani river basin of Tamil Nadu, India and kept in cement tanks. Three inducing hormones viz, Ovaprim, Ovatide and WOVA-FH were injected at the rate of 0.5, 1.0, 1.5 and 2.0 ml/kg body weight in order to induce oocyte maturation and ovulation. After 10-13h of injection at a water temperature of 27±-0.5°C, stripping of eggs and in vitro fertilization was done. Ovaprim gave maximum (94.67%) hatching rate followed by Ovatide (90.33%) and WOVA-FH (77.33%).

Preliminary success on hormone induced captive breeding of goldspot mullet, Liza parsia (Ham.)

An attempt was made to breed goldspot mullet, Liza parsia in captivity through hormone induction. The fish started spawning 35-36 hours after a single dose of 2ml ova prim per kg body weight. Hatching of fertilized eggs completed within 42-48 hours after spawning. The mean hatching rate (%) was 71.33±12 corresponding to the fertilization rate (%) of 64±12. The larvae started its first external feeding on the third day and attained a length 2.5±0.25 mm. The salinity of both breeding and rearing cisterns was 20‰ and temperature was maintained at 22-23°C.

Assessment of impact of ten days fishing ban in the major spawning grounds of hilsa (Tenualosa ilisha, Fisher and Bianchi, 1984)

A study was undertaken to find out the impact of ten days fishing ban in the major spawning grounds of hilsa during October to December. The study revealed a positive impact of fishing ban during spawning season on reproduction success of hilsa. Proportionate distribution of male and female hilsa also showed a significant level of distribution in and around the spawning grounds where the ratio was 35:65. Percent of oozing hilsa during fishing ban period in the spawning grounds was observed 1.61%. About 5% spent hilsa was observed in the fish landing centers and was compared with the data of Global Environment Facility (GEF) and Bangladesh Fisheries Research Institute (BFRI) studies and was found about 2.80-3.57 times higher than that of the findings of 2002 and 2003. Estimated egg production value showed about 46,800 Kg of eggs could have been produced that indicating a positive impact of 10 days fishing ban in spawning season. Abundance of higher percentages of gravid hilsa was found which were not available in the same quantity and condition in the non-fishing ban period. During the present investigation, fairly higher amount of spent hilsa and juveniles were also observed in the spawning grounds. Along with the jatka fry, spawn and fries of other fishes were also found in higher quantity than the previous years and thus it is assumed that fishing ban also might have positive impact on the successful breeding of other fishes. Overall, the fishing ban was found effective for successful breeding of hilsa.

Effects of dietary vitamin E on the growth and breeding performance of Ompok pabda

A 90 day feeding trail was conducted to investigate the effect of vitamin E on the growth and breeding performance of Ompok pabda. A total of 84 healthy female brood fish (41.10±0.44g) were divided into 4 treatments i.e. treatment T1 T2, T3 and T4 having three replications each. The fish were fed twice a day with a standard feed (40% protein) having 4 doses of vitamin E viz. 0 (served as control), 50, 100 and 150mg vitamin E/kg feed. At the end of the feeding trial, the brood fish were induced to breed with PG extract to observe the effect of vitamin Eon feed. After rearing for 90 days with the experimental feeds, it was found that weight gain and specific growth rate of brood fish fed with 100mg vitamin E/kg feed (treatment T3) was the highest (14.78±0.38g and 2.99±0.11) while 150mg vitamin E/kg feed (treatment T4) fed fish gave the poorest result (2.97±0.89g and 1.21±0.32). There was no significant difference in terms of length gain of brood fish among the different treatments. The brood fish were induced to breed with equal dose of PG extract (18 and 12mg PG/kg body weight for female and male respectively) to observe the dietary effect of vitamin E on breeding performance. The highest ovulation, fertilization and hatching rate of eggs were found to be 81.48±6.41%, 84.04±3.53% and 68.59±5.03% respectively in the brood fish of treatment T1 while the poorest (33.33±00%, 52.35±5.02% and 45.70±7.24% respectively) were found in the brood fish under treatment T4. The results suggest that inclusion of 100mg vitamin E/kg feed is best for enhancing the breeding performance of O. pabda brood fish indicating that vitamin E content has a positive impact on reproduction of fish. The present results also imply that inclusion of higher level of vitamin E exerts an antagonistic effect in terms of growth and breeding performance of this species.

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.

Determination of some dominant genetic characteristics and morpho-meristic features for selection of rainbow trout (Oncorhynchus mykiss) broodstocks

In this study, Iranian and French male and female Oncorhynchus mykiss broodstocks were divided into two groups 50 and 24 respectively in Research center of genetic and breeding of coldwater fishes, Yasouj, Iran and the genetic structure of them was investigated using 6 microsatellite markers. Then 19 morphometric and 5 meristic of broodstock were measured and compared in two populations. Along with broodstock maturation, fertilization 1:1(female:male) were randomly assigned and occurred in 25 of 12 Iranian and French treatment respectively. Reproductive parameters were recorded for the whole family. Average number of observed alleles in Iranian and French stocks was 6.68 and 6.83, respectively. Average number of effective alleles in Iranian and French stocks was 3.13 and 3.45 respectively. Fixation index Fst was calculated based on allelic frequency between two stocks was 0.058 with significant difference between 2 stocks. Morphometric analysis showed significant difference between two stocks in 8 characteristics. Meristic characters was without significant difference in broodstock groups. Eyed percentage for french broodstock calculated zero and deleted. Fertilization rate (100-0), the eyed percentage (98- 0), The hatch rate (98-0), the average fecundity 4114.708, the average eggs size 4.88 mm, Survival in the first three months 19-73% calculated for Iranian broodstocks. Considering the quality of eggs and larvae at different stages and selection between the different family and the within family remained 10 treatments and are kept as future broodstocks. The relationship between fecundity - egg size, fecundity - weight , fecundity - length, egg size- weight was performed using regression. The results showed that Fecundity was influenced more by weight and productive length. The research is beginning to ID the broodstock in our country.

Study on macroscopic, microscopic and hormonal changes in gonads of the crayfish Astacus leptodactylus in Aras Dam

This study was carried out to seasonal determination of some morphological characteristics, Seasonal fecundity, Seasonal fluctuations of vertebrate-type steroids and seasonal analysis of gonadal histology in both female and male sexes of freshwater crayfish (Astacus leptodactylus Eschscholtz 1823) in the area of Aras dam Lake. Crayfish were collected respectively in June, August, November (2011) and January (2012). The average length and weight of male crayfish was higher than that of females. %GSI of females fluctuated within an extended range (between 0.6 and 13.5% from June to January). Both of synchronous and asynchronous ovaries were seen in August sampled ovaries; however asynchronous form was higher than another. The annual reproductive cycle of male A. leptodactylus was surveyed by study on the seasonal changes of the external appearance of the testes and vasa deferentia, fluctuations in the gonadosomatic index (GSI%) and the histological analysis of the male reproductive system. Based on the histological differentiation of testis, spermatogenisis devided to 5 separated stages. The findings suggested asynchronous testis in the species A.leptodactylus. The presence of primary spermatophore layer may help keeping spermatozoa alive while the secondary spermatophore layer may produces spermatophore or synthesize of acellular material which forms spermatophre. Pleopodal fecundity was 37.3%lower than ovarian fecundity observed. The significantly higher number of eggs attached to 3rd and 4th pairs of pleopods. The egg number and gonadosomatic index increased with female size while egg weight and egg diameter didn’t increase with female size. Hemolymph levels of 17β-estradiol and progesterone followed a similar fluctuation pattern with % GSI in females, while testosterone didn’t follow the mentioned pattern. The testis of November sampled crayfish presented significantly higher gonadosomatic (%GSI) index (P < 0.05).The most observed gonadosomaticindices were 13.5%(forfemales) and 1.21% (for males, in autumn. Althogh the lowest GSI was (0.50%) formales in spring and (0.26%0 for spent females in January. Testosterone which followed a similar pattern with %GSI in males increased remarkably in November. 17β-estradiol increased strictly in January. The strictly enhancement of the three estroid hormones in January in both male and female sexes could bedue totheir stimulating role in in spermatophre and egg lying in the mating season (In January). Most of the ovaries followed the asynchoronous growth pattern. Also the testes presented asynchoronous growth pattern in autumn.