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.

Demonstration of some selected aquaculture technologies under farming system research in Jessore and Santahar, Bangladesh

BFRI evolved some selected aquaculture technologies viz. polyculture of carps in perennial ponds, monoculture of short cycled fish species (BFRI super strain) in seasonal ponds and prawn seed production through backyard hatchery system have been demonstrated under Farming System Research (FSR) component in Jessore and Santahar regions. Both polyculture of carps and monoculture of short cycled fish species technologies were tested in farmer's ponds in Kaium Kula village near Jessore town. In polyculture trials, seven species comprising of silver carp (Hypophthalmichthys molirrix), catla (Catla catla), rohu (Labeo rohita), grass carp (Ctenopharyngodon idellus), common carp (Cyprinus carpio), mrigal (Cirrhinus cirrhosus) and silver barb (Barbonymus gonionotus) were stocked @ 9,500 (ratio 6:2:4:2:1:5:5); 10,750 (ratio 6:2:4:2:1:5:5) and 12,000 (ratio 6:2:4:2:1:5:4) fish/ha respectively in ponds of T1, T2 and T3 having three replications of each. The mean highest fish production was 3,148 kg/ha in T3, followed by 2,899 kg/ha in T1 and 2,875 kg/ha in T2. Production of T3 was significantly different (P<0.05) than both T1 and T2, while there was no significant differences (P>0.05) between the production of T1 and T2. In case of trial of short cycled fish species, two treatments were tested: T1 (comprising of BFRI super strain of Nile tilapia, silver carp, common carp and silver barb; ratio 3:5:1:1) and T2 (having only BFRI super strain of Nile tilapia). Stocking density in both the treatments were same (20,000 fish/ha). In this trial average production was higher in T1 (2,743 kg/ha) than that of T2 (2,369 kg/ha) but the production figure in these two treatments was not significantly different (P>0.05). Demonstration of backyard prawn hatchery technology was tested at Santahar region of Bogra district, North-west part of Bangladesh. This hatchery consisted of three main components i) bio-filter, ii) rearing tank unit (chari) and iii) air blower/air pump unit. Plastic drum of 200-250 l capacity and cemented chari of 200-250 l capacity were used as bio-filter and larval rearing containers respectively. A 0.5 hp air blower with 6 aquarium air pump were used to operate the aeration system in the hatchery. Diluted sea water (10-12 ppt) made from brine solution (200-250 ppt) collected from salt-bed was used in the backyard hatchery system of hatching of eggs and rearing of larvae. Rearing of first stage zoea-larvae was reared in three rearing tanks following the stocking densities of 40, 50 and 60/l of water respectively. Production of post-larvae were 20±0.82, 22±1.12 and 28±1.63/liter of water in treatments I, II and III respectively in 38, 40 and 39 days rearing period.

Identification and culture of common diatoms as possible feed for Penaeus monodon

Diatoms were collected from Buyuan Bay, and from the hatchery tanks at Tigbauan, to determine the commonly occurring species, the feasibility of culturing these species, and the potential of these selected species as food for larval P. monodon. The commonly occurring diatoms were identified as Chaetoceros calcitrans, Navicula grimmei, Nitzchia seriata, Nitzchia closterium and Amphiprora sp. These diatoms were isolated and unialgal cultures prepared. Protein content analysis using the micro-Kjildahl method gave the following result: C. calcitrans, 11 . 78%; Nitzchia seriata, 25%; Nitzchia closterium, 30 . 5%; Navicula grimmei, 9 . 06% and Amphiprora sp. 8 . 96%. Feeding experiments were conducted to determine acceptability of the different diatom species and percentage survival of larval stages Z SUB-1 -M SUB-2 . Larvae were placed in 4-l capacity plastic containers with a stocking density of 10/l. The results of several feeding trials using the different mass-produced diatoms are summarized. From the data gathered, C. calcitrans appears to be the most promising candidate as feed for zoea and mysis stages of P. monodon. The average percentage survival of C. calcitrans was 63 . 76% for the 3 trials, and as high as 82 . 22% in the third trial. Comparatively high percentage survival of larvae was also recorded when Nitzchia seriata (48 . 17%) and Nitzchia closterium (67 . 6%) were given as feed, while both Amphiprora sp. and Navicula grimmei gave 0% survival. The poor results with Amphiprora sp. and Navicula grimmei may be due to their low protein content (8 . 96% and 9 . 06%, respectively) and the inability of the larvae to ingest them. Navicula and Amphiprora were observed to cling to the appendages of the larvae and to settle down in the medium making them unavailable to the larvae. Low survival was also noted when frozen C. calcitrans was used (14 . 25%). This may be due partly to the effect of the floculating agent (ALSO SUB-4 . 25 g/l) used in concentrating the diatoms. When protein contents of C. calcitrans, N. seriata and N. closterium are compared, the 2 Nitzchia species have relatively higher protein contents than C. calcitrans and, therefore, could be the more desirable feed candidates. However, few feeding trials were made using Nitzchia so that additional investigations will have to be done on this aspect.

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.

Effects of furanace on Brachionus

Tiger prawn P.monodon) larvae utilize Brachionus a rotifer, as food in the Zoea 3 and mysis stages when they change from an herbivorous to an omnivorous diet. The present work aims to show the effects of furanace on the population growth of Brachionus. Cultures of Brachionus were obtained and fed with Chlorella at a density of 1-2x10 SUP-6 cells/ml. Five liters of the culture water were placed in each of 4 white, circular, 152x304 mm plastic basins. The mean initial densities of the rotifer ranged from 26 . 5 to 38 . 5 individuals/ml. The concentrations of furanace were 0, 1, 2 and 3 mg /l. The cultures were vigorously aerated. Population growth was observed after 3, 6, and 9 hours of exposure. The cultures were thoroughly mixed before samples were taken to ensure an almost equal distribution of the rotifers in the water. To facilitate the counting of the rotifer, one drop of Lugol's solution was added to each sample. This immobilizes the rotifer as well as stops further reproduction. Individuals with only the lorica left or with badly deformed lorica were considered dead. Population counts were done using a Sedgwick-Rafter counting chamber. Among the different durations of exposure, the percentage survival of the populations in the furanace baths were highest after 3 hr. There were slight increases in the control and 2 mg/l and slight decreases in 1 and 3 mg/l. The differences in the mean densities are statistically insignificant at . 01 significance level. After a 6-hr exposure, the control population reached its peak density with a survival of 89%. Populations in furanace baths decreased to 88 . 5% in both 2 and 3 mg /l followed closely by 87% in 1 mg/l. Again, no statistical differences exist among all the levels. The mean percentage survival in 1 and 2 mg/l increased (89% and 91%, respectively) after a 9-hr expsoure, while those in the control and 3 mg/l decreased to 86 . 5% and 88 . 25%, respectively. There were no marked differences in appearance noted among the individuals in furanace baths and those in the control.

Use of fermented kitchen waste in rearing Penaeus monodon larvae

Fermented vegetable and kitchen wastes are available as feeds for not only zoea but also mysis and up to certain points in the postlarval stages of sugpo, Penaeus monodon. It is recommended that the hatchery use fermented wastes as larval feed for P.monodon when diatoms or brine shrimp nauplii are lacking or in short supply. Among three stages namely, zoea, mysis and postlarva, the survival rate during postlarva particularly after P SUB-4 was quite low. The problems encountered are as follows: (a) how to prevent fermented particles from lumping, (b) how to prevent them from easily sinking to the bottom, and, (c) how to prevent bacteria and fungi, particularly Lagenidium sp blooming.

Effect of nitrite, ammonia, and temperature on P. monodon larvae

P. monodon larvae were studied for the effects of temperature, ammonia, and nitrite on survival. Toxicity levels of nitrite were found to vary with larval stage. Larvae could tolerate ammonia up to about 10 ppm, with the effect more clearly shown by the zoea stage. Survival and growth were not significantly affected by temperature, although moulting was enhanced at temperatures higher than 29 C. Larvae of P. monodon have lower tolerance toward nitrite and ammonia compared to postlarvae. Although high survival was obtained at low levels of nitrite and ammonia, it is still necessary to know their effects on metabolism, in order to examine possible biochemical parameters for diagnosing sublethal toxicity or stress.

Preserved algae as food for Penaeus monodon larvae

Uni-algal cultures of C. calcitrans, S. costatum, T. chui and Isochrysis sp. obtained from the laboratory were harvested. The harvest, preserved by either freezing or sun-drying, was fed to the larvae of P. monodon . Among the test algal species, Chaetoceros and Tetraselmis were used in larval feeding trials with frozen food while Chaetoceros, Tetraselmis and Isochrysis species were utilized in feeding experiments with sun-dried algae. Their relative effects on larval survival and development were assessed. Results showed that, except the alum-flocculated cells, both frozen Chaetoceros and Tetraselmis can support survival at the zoea stage. Best survival of 68% was atained with dried Chaetoceros followed by Tetraselmis at 44%. Dried Isochrysis did not perform as well, a significantly low survival of only 25% was obtained.

Growth and survival of Penaeus monodon Fabricius zoeae on different diatom feeds

A study was undertaken to determine the most suitable food for Penaeus monodon zoea out of mixed diatoms, Skeletonema costatum, Chaetoceros calcitrans and C. gracilis. Results indicate that mixed diatoms and C. gracilis are the better food. Addition of zooplankton, e.g.rotifers, is also recommended as early as Z3 since the larvae start to be omnivorous at this stage.

The study of Indian shrimp larvae feeding by enriched rotifer with vitamin C and unsaturated fatty acid (epa, dha) and its effects on growth factors, survival rate and environmental stresses

In this experiment, the feeding of Indian white shrimp larvae by unenriched rotifers (treatment 1) and enriched with highly unsaturated fatty acid (treatment 2) and highly unsaturated fatty acid along with vitamin C (treatment 3) on the growth factors, survival and resistance against salinity and formalin stress tests were studied and their differences with control treatment including newly hatched Artemia nauplii is compared. In this the study four treatments in a completely randomized design with 3 replicates per treatment were used. Farming of shrimp larvae of Zoea II to postlarvae 5 was done in 20 liter plastic bucket. Present results indicated that growth factors and survival rate of stage Zoea II to postlarvae 1 in treatments 1, 2 and 3 improve rather than control in which this case was due to optimal size rotifer rather than Artemia nauplii. Also, treatments 2 and 3 feeding with oil liver cod emulsion enriched rotifer have the highest concentration of DHA (mg/g DW) and the ratio DHA/EPA in which due to have shown the highest growth factors and a significant difference (P<0.05) with treatments 1 and control. The highest survival at stage PL1 were observed in treatment 3 that was enriched with ascorbyl palmitate in which have to the synergistic properties of vitamin C rather than treatments 2, 1 and control and showed a significant difference (P<0.05). But in stage PL5 the highest amount of growth and survival rates were related to control treatment which showed a significant difference (P<0.05) with other treatments that control has higher size rather than treatments 1, 2 and 3. Also, among experiment treatments that the two treatments 2 and 3 due to enrichment had higher growth and survival rates compared with treatment 1 in which their differences have also been significant (P<0.05). In the case of stress tests, results indicated that the highest survival rate has been reported when specimens were offered a diet containing high levels of highly unsaturated fatty acids with vitamin C. So that in stage PL1 in the salinity stress tests 10 and 20 ppt the highest survival rate was observed in treatment 3. As for the second, treatment 2 showed a significant difference (P<0.05) with treatment 3. It is worth mentioning that treatment 3 showed a higher survival rate compared to treatment 2 due to the synergistic properties of vitamin C. The difference between these two treatments with treatment 1 and control was also significant. No significant difference was observed in formalin stress test 100 ppm in this stage between treatments 3 and 2 which shows the highest survival rate. But their difference with treatments 1 and control was significant (P<0.05). Also, in stage PL5 in the salinity stress tests 10 and 20 ppt the highest survival rate was observed in treatment 3 which showed no significant difference (P<0.05) with control treatment. While their difference in the amount of survival rate with treatment 1 and 2 was significant (P<0.05). In this stage, the highest observed survival rate in formalin stress test 100 ppm included treatments control, 3 and 2 among which there were no significant differences (P<0.05). While the difference between these three treatments with treatment 1 was significant.

Improvement of polyploidy induction in Eriocheir sinensis

An incubating temperature of 15 degreesC is used to induce triploidy in Etiocheir sinensis through inhibition of the release of polar body H, and that of 18 degreesC to induce tetraploidy through inhibition of the first cleavage. Flow cytometry is used to identify the ploidy in different developmental stages. For induction of triploidy in fertilized eggs in vitro, the highest induction rate observed in blastula by cytochalasin B, 6-DMAP and KCI is 49.1%, 51.7% and 77.5%, respectively. In the KCI treatment of pregnant crabs with the fertilized eggs, the highest triploid induction rate observed in the zoea is 85.3%. For induction of tetraploidy, the highest induction rate observed in the blastula by cytochaslasin 13, 6-DMAP and KCI is 50.3%, 54.9% and 79.8% respectively. In the KCI treatment of pregnant crabs with the fertilized eggs, the highest induction rate in zoea is 27.3%. Through this study such difficulty as in vitro culture is overcome. Triploid zoea Etiocheir sinensis has been developed for the first time. The induction rate of tetraploid zoea has also been greatly improved.

Foreign gene transfer into Chinese shrimps (Penaeus chinensis) with gene gun

Plasmids pG DNA-RZ1 with a GFP (green fluorescent protein) reporter gene and a ribozyme gene incising penaeid white spot baculovirus (WSBV) were first introduced into the fertilized eggs of Chinese shrimps by gene gun. The treated and control samples of different development stages were observed with a fluorescent microscope. The transient expression of GFP gene was high in nauplius and zoea larvae. Results from RT-PCR and PCR for adults showed that the foreign genes had been transferred into the shrimps and had expressed the corresponding proteins. This work has established a transgenic method for penaeid shrimps, which will set base for the application of genetic engineering breeding into industry.

A Dicer-1 gene from white shrimp Litopenaeus vannamei: Expression pattern in the processes of immune response and larval development

Dicer is a member of the RNAase III family which catalyzes the cleavage of double-stranded RNA to small interfering RNAs and micro RNAs, and then directs sequence-specific gene silencing. In this paper, the full-length cDNA of Dicer-1 was cloned from white shrimp Litopenaeus vannamei (designated as LvDcr1). It was of 7636 bp, including a poly A tail, a 5' UTR of 136 bp, a 3' UTR of 78 bp, and an open reading frame (ORF) of 7422 bp encoding a putative protein of 2473 amino acids. The predicted amino acid sequence comprised all recognized functional domains found in other Dicer-1 homologues and showed the highest (97.7%) similarity to the Dicer-1 from tiger shrimp Penaeus mondon. Quantitative real-time PCR was employed to investigate the tissue distribution of LvDcr1 mRNA, and its expression in shrimps under virus challenge and larvae at different developmental stages. The LvDcr1 mRNA could be detected in all examined tissues with the highest expression level in hemocyte, and was up-regulated in hemocytes and gills after virus injection. These results indicated that LvDcr1 was involved in antiviral defense in adult shrimp. During the developmental stages from fertilized egg to postlarva VII, LvDcr1 was constitutively expressed at all examined development stages, but the expression level varied significantly. The highest expression level was observed in fertilized eggs and followed a decrease from fertilized egg to nauplius I stage. Then, the higher levels of expression were detected at nauplius V and postlarva stages. LvDcr1 expression regularly increased at the upper phase of nauplius, zoea and mysis stages than their prophase. The different expression of LvDcr1 in the larval stages could provide clues for understanding the early innate immunity in the process of shrimp larval development. (C) 2010 Elsevier Ltd. All rights reserved.

Digestion of natural food by larval and post-larval turbot Scophthalmus maximus

The digestion of natural, mainly crustacean zooplankton, by different age groups of turbot Scophthalmus maximus larvae was evaluated by comparisons of visual appearance, dry weight and carbon and nitrogen content of fresh food organisms with material recovered from faeces. Visually, the degree of digestion of food particles ranged from no discernible change of lamellibranch larvae, copepod eggs, intact copepod faecal pellets and some phytoplankton species, to varying degrees of removal of body constituents in copepods, cladocerans and decapod zoea. For crustaceans, the proportion of body constituents removed was related to the size and construction of their apparently indigestible exoskeleton. Uppon defaecation larger organisms showed the greatest percentage loss in dry weight and carbon. A high percentage of nitrogen was extracted from all organisms.

Feeding rates and prey selectivity of planktonic decapod larvae in the Western English Channel

Meroplankton are seasonally important contributors to the zooplankton, particularly at inshore sites, yet their feeding ecology is poorly known relative to holoplankton. While several studies have measured feeding in decapod larvae, few studies have examined the feeding rates of decapod larvae on natural prey assemblages throughout the reproductive season. We conducted 8 feeding experiments with Necora puber, Liocarcinus spp. and Upogebia spp. zoea larvae collected from the L4 monitoring site off Plymouth (50°15.00′N, 4°13.02′W) during spring–summer 2009 and 2010. This period spanned moderate-to-high food availability (0.5–1.6 µg chl-a L−1), but a great range in food composition with small cells <20 µm dominating in 2010. Daily rations averaged 17, 60 and 22 % of body C for the 3 respective decapod species. Clearance rates differed according to prey type, and all 3 decapod genera showed evidence of selection of dinoflagellates. Importantly, small cells including nano- and pico-plankton were ingested, this being demonstrated independently by flow cytometric analysis of the feeding experiments and molecular analysis. PCR-based analysis of the haptophyte portion of the diet revealed ingestion of Isochrysis galbana by decapod larvae in the bottle incubations and Isochrysis galbana and Phaeocystis globosa by decapod larvae collected directly from the field. This study has shown that pico- and nano-sized plankton form an important supplement to the diverse and variable diet of decapod larvae.