生殖激素对奶水牛卵母细胞成熟及发育潜能的影响

本试验以屠宰场获取的奶水牛卵巢为试验材料,收集卵母细胞进行体外成熟培养(IVM)、体外受精(IVf)及早期胚胎培养(IVC).研究激素(FSH、LH、E2、P4)的不同浓度对奶水牛卵母细胞成熟和早期胚胎发育的影响,以期探讨奶水牛卵母细胞成熟和早期胚胎体外培养发育机制,优选不同激素的最佳浓度.结果表明:添加FSH试验组奶水牛颗粒细胞扩散率和卵裂率高于未添加试验组(P<0.05);添加LH试验组奶水牛颗粒细胞扩散率、卵裂率及8-细胞率与未添加试验组比较,差异不明显(P>0.05);17β-E2试验组(1.0μg/mL)的奶水牛颗粒细胞扩散率、卵裂率及8-细胞率高于未添加试验组(P<0.05);添加P4试验各组(0.9μg/mL、1.2μg/mL)的颗粒细胞扩散率明显低于未添加试验组(P<0.01).

Laboratory culture of Moina with organic ana inorganic fertilizers

An experiment was conducted in the laboratory condition to determine the effect of organic (poultry drop, cow dung and mustard oil cake) and inorganic fertilizer (urea) on production, reproduction rate and maturation time of Moina species. Production rate was also determined in both aerated and non-aerated system in plastic containers with carrying capacity of 2.5-liter each. Total production was significantly higher in both aerated (2475 individuals/2.5 l water) and non-aerated (3253 individuals/2.5 l water) containers using poultry manure compared to other fertilizers. Moreover, the reproduction rate and maturation time in poultry drops showed distinct efficacy in Moina species. Reproduction rate of 11 individuals was the maximal while lowest maturation time was found 78 hours. Reproduction and maturation were induced surprisingly in test tube where the aeration system was absent.

Emigration dynamics of three species of penaeid prawn from backwaters and tidal ponds of Cochin, India

Emigration of three species of penaeid prawn from backwaters and tidal ponds were studied. Considerable diel, tidal, lunar and seasonal fluctuations were observed in emigration process, which was almost nocturnal. Rate of emigration and composition of emigrants varied with time of migration. Large pulses of emigration always coincided with spring tides with major peak during new moon. Seasonal variation was observed with peak emigration of Penaeus indicus during monsoon months and that of Metapenaeus dobsoni and Metapenaeus monoceros during pre-monsoon. Rate of emigration was relatively large from shallow tidal ponds. It correlated directly with the prevailing environmental conditions and juvenile density. Instantaneous rate of emigration was also large in seasonal ponds. The basic stimulus for emigration is the urge for sexual maturation. Coupled with it ecological changes in the habitat have been causing various patterns in migration.

Suggestive role of eye-stalk of Macrobrachium gangeticum Bate, 1868 in regulation of its reproductive behaviour

Role of eye-stalk of Macrobrachium gangeticum Bate, 1868 in its reproductive behaviour has been examined by conducting deletion and addition experiments. Eye-stalk ablation induced gonadal maturity in both sexes, leading to change in colour and size of ovaries and increase in GSI and oocyte diameter in females and increased length of testes and diameters of seminiferous tubules in males. Injection of eye-stalk extracts tended to at least partly restrict the effects in both sexes. The experiments thus suggested that the eye-stalk of M. gangeticum released some gonad inhibiting factors.

Seed production and culture techniques of Genetically Improved Farmed Tilapia (GIFT) in brackishwater environment

Effects of different levels of salinity on survival, growth and gonadal development of Genetically Improved Farmed Tilapia (GIFT) were studied under laboratory conditions in glass aquarium, for a period of ten weeks. The initial individual size of the GIFT was 20.23±4.45 and the salinity levels tested were 0, 5, 10, 15 and 20 ppt. The highest survival of 87.5% was found in 0 ppt and the lowest 60.5% in 20 ppt. Though the survival decreased progressively with increased salinity, there were no significant differences (P>0.05) among 0, 5, and 10 ppt. Similar to what has been observed in survival, the specific growth rate (SGR %/day) also decreased as of 1.30, 1.24, 1.08, 0.90 and 0.71, respectively, with the increased salinity of 0, 5, 10, 15 and 20 ppt. The gonadal development was highest in 0 ppt with a GSI value of 3.75 and lowest of 2.01 in 20 ppt. In the second experiment, gonadal development and seed production performance of GIFT in brackishwater condition were investigated for a period of three months. Each of the three fine meshed hapas of 20 square meters made from nylon net was placed in a freshwater (0 ppt) and in a brackish water (10-15 ppt) pond of the Brackishwater Station (BS). GIFT of 65 g average weight from a single cohort were stocked into three hapas at a rate of 2 per m. The male vs female ratio was 1:3. The development of gonad was faster with the higher gonadosomatic index (GSI %) of 3.85 % in freshwater condition than that of 2.73 % in brackish water. Within three months of the study period, a total of 70,510 and 44,250 GIFT fry were produced respectively, in freshwater and brackishwater conditions. Finally under third experiment, a participatory on-farm trial was carried out to evaluate the production performance of GIFT in monoculture and in polyculture with silver barb in coastal freshwater pond conditions. Nine ponds were selected for three treatment combinations of GIFT monoculture (T1), GIFT and silver barb polyculture (T2), and silver barb monoculture (T3). The ponds have been stocked in April, 05 at a density of 25,000 fry per ha. Fishes were fed with rice bran at the rate of 6% bw per day. In one month culture period, GIFT attained an average weight of 16.27 g in monoculture and 17.23 g in polyculture, against an average stocking weight of 0.37 g. Silver barb reached an average weight of 16.62 g in polyculture with GIFT and 10.01 g in monoculture, against an average stocking weight of 3.79 g.

An ARGONAUTE4-containing nuclear processing center colocalized with Cajal bodies in Arabidopsis thaliana.

ARGONAUTE4 (AGO4) and RNA polymerase IV (Pol IV) are required for DNA methylation guided by 24 nucleotide small interfering RNAs (siRNAs) in Arabidopsis thaliana. Here we show that AGO4 localizes to nucleolus-associated bodies along with the Pol IV subunit NRPD1b; the small nuclear RNA (snRNA) binding protein SmD3; and two markers of Cajal bodies, trimethylguanosine-capped snRNAs and the U2 snRNA binding protein U2B''. AGO4 interacts with the C-terminal domain of NRPD1b, and AGO4 protein stability depends on upstream factors that synthesize siRNAs. AGO4 is also found, along with the DNA methyltransferase DRM2, throughout the nucleus at presumed DNA methylation target sites. Cajal bodies are conserved sites for the maturation of ribonucleoprotein complexes. Our results suggest a function for Cajal bodies as a center for the assembly of an AGO4/NRPD1b/siRNA complex, facilitating its function in RNA-directed gene silencing at target loci.

Dynamic regulation of ARGONAUTE4 within multiple nuclear bodies in Arabidopsis thaliana.

DNA methylation directed by 24-nucleotide small RNAs involves the small RNA-binding protein ARGONAUTE4 (AGO4), and it was previously shown that AGO4 localizes to nucleolus-adjacent Cajal bodies, sites of snRNP complex maturation. Here we demonstrate that AGO4 also localizes to a second class of nuclear bodies, called AB-bodies, which are found immediately adjacent to condensed 45S ribosomal DNA (rDNA) sequences. AB-bodies also contain other proteins involved in RNA-directed DNA methylation including NRPD1b (a subunit of the RNA Polymerase IV complex, RNA PolIV), NRPD2 (a second subunit of this complex), and the DNA methyltransferase DRM2. These two classes of AGO4 bodies are structurally independent--disruption of one class does not affect the other--suggesting a dynamic regulation of AGO4 within two distinct nuclear compartments in Arabidopsis. Abolishing Cajal body formation in a coilin mutant reduced overall AGO4 protein levels, and coilin dicer-like3 double mutants showed a small decrease in DNA methylation beyond that seen in dicer-like3 single mutants, suggesting that Cajal bodies are required for a fully functioning DNA methylation system in Arabidopsis.

In vitro construction of a recombinant human embryonic brain-derived neurotrophin-4 gene and pEGFP-N1 vector

BACKGROUND: Neurotrophin-4 (NT-4) can promote neuronal growth, development, differentiation, maturation, and survival. NT-4 can also improve recovery and regeneration of injured neurons, but cannot pass through the blood-brain barrier, which limits its ac

A simple method of tagging prawns

The recognition of individual animals is crucial to many aspects of research. Prawns (Penaeus monodon) present unique difficulties in this respect since they molt regularly. Thus, almost all tagging and marking methods developed for prawns so far have proven inadequate. Some tags or marks are lost during molting; others cause injury to the prawns. A new and efficient method has been developed at the Igang Seafarming Station of the Aquaculture Department. Rectangular brass tags measuring 5 mm by 20 mm and numbered consecutively are used. The prawn is held gently but firmly at the base of the carapace with the left hand while the right hand slips the brass tag around the stalk of the unablated eye and presses the tag gently. All tagging must be made under water to avoid stress. From May 29 to September 7 1977 to a total of 348 unilaterally-ablated adult female prawns were tagged on the unablated eyestalk in 5 batches to enable individual observations on gonadal maturation, molting, and growth. Periodic examinations were made four times a month to coincide with the different phases of the lunar cycle. On each examination, survival and recovery rates were recorded. The data included death due to immediate mortality during ablation and loss to cannibalism for the duration of the experiments. In all five tagging experiments, most of the prawns recovered had their tags intact. These included even dead and molting animals. The eyestalk tagging method is suitable for prawns because the tags can be attached without causing injury and has no effect on the rate of growth, maturity, molting and behavior of the animal. The tags are identifiable and permanent; they remain attached to the animal even after death.

Biology and farming of the green mussel Mytilus smaragdinus

Biological investigations were carried out in Sapian Bay, Capiz from November 1975 to December 1976 with samplings conducted fortnightly. Histological studies on the gonad reveal a high percentage of ripe and spent females during the month of April and May, and ripe to near ripe during November to December. However, larval counts were highest on February 25, 1976 with 253 mytilid larvae per haul compared to 0-79 per haul during all other months. The high larval count was followed by the highest spat settlement during the next sampling period two weeks later, with the spat collector set in the water during the February 25 sampling. The four materials tested, blue polypropylene fiber rope, black polypropylene fiber, and coir rope, all had their highest spat counts during this period with an average of 471 spats per standard 10 cm rope piece. The range during the other time periods is 2-283 spats. Of the 4 materials tested, the black fibrillated polypropylene film had the highest larval counts in 15 out of a total of 25 sampling periods. The blue rope was the poorest spat collector. Coconut husk was tested later on and it proved to have a very high catchability, with spats completely enveloping the husk surface. Growth monitored from one cohort in Sapian Bay averaged 10 mm per month. 50-60 mm is considered marketable size. Trial growth experiments with transplanted mussels were also conducted at Igang Bay in Guimaras Island, Makato River in Aklan, and a milkfish pond in Leganes, Iloilo. Survival in Igang was less than 50% after the second week, and the condition of the surviving mussels can be described only as 'watery' with the mantle completely transparent. Mortality was minimal in Makato but the growth rate was only 30% that of Sapian Bay. The pond experiments were terminated due to severe crab predation.

The propagation of the mud crab Scylla serrata (F.) de Haan

The mud crab Scylla serrata is an important commercial species found in many brackish areas in the Philippines. During spawning and hatching, the berried females migrate to the sea. Seeds for pond stocking are obtained from the wild. Because of the unpredictability of seed supply, there is a need to propagate the species artificially. Thus, spawning, larval rearing, maturation, and rematuration of the species are being studied. The first attempts at hatching S. serrata were successful with rates varying between 75% and 90%. Two out of three trials on larval rearing yielded a few megalops. The first zoeal stages were fed diatoms, rotifers, Artemia salina, and bread yeast. Overfeeding programs were implemented during the critical premolting periods to prevent weakening of the larvae and lessen cannibalism. Larval weakening during the premolt makes them susceptible to attacks by fungi like Lagenidium and ciliates like Vorticella. S. serrata larvae survived salinity levels as low as 15 ppt until the 14th day of rearing. Other larvae were able to survive in salinities of 30-32 ppt for 8 to 13 days. Zoeal molting was hastened by lowering the salinity to 25-27 ppt. Artificial broodstocking of juveniles and adult crabs has been made possible using a simple refuge system made of three-compartmented hollow blocks. This system has been helpful in minimizing fighting among crabs. Remarkable growth rates have been observed with feeds like mussel meat and trash fish. Average growth increments of 11 mm carapace length and 20 . 35 g body weight have been observed every fortnight. A newly spent spawner could gain additional weight of 22 . 5 g in only 6 days. Feeding rates of juveniles and adult crabs have been established based on the average body weight from an experiment using mussel meat. Crabs feed more at night. In another experiment, eyestalk ablation was found to be effective in inducing growth and mating. Aside from hastening the molting process, copulation is induced even among the small crabs (average carapace length = 55 mm). Natural mating lasts about 26 hr. A copulation which lasted for seven days with a break in between was observed.

Notes on ovarian rematuration of ablated sugpo (prawn) Penaeus monodon Fabricius

To what extent spent P. monodon females can remature and spawn successive broods is an important question in terms of recycling spawners in a commercially viable operation. Corollary to this is the quantity and quality of fry from rematured females in comparison to those from first spawning. Of 347 experimental females, only 10.1% had a second spawning, and 1.4% a third spawning. To a large degree the low rate of rematuration is due to high spawner mortality - average survival period after spawning was only 6 days in a sample of 176 spawners. It took an average of 23 days after ablation for a prawn with undeveloped ovaries to mature and spawn. An ablated female may have another spawning in as little as 5 days after the previous one. Average fecundity was 180,000 eggs per second spawning, and 140,000 eggs per third spawning. The average number of eggs from first spawning ablated females was 110-120,000. Hatching rate was lower for rematuration: 44% for second spawnings, and 35% for third spawnings, as compared to 64% for first maturation.

Viability of Penaeus monodon eggs after simulated transport conditions

P. monodon spawners, transported from maturation pens suffer from stress which in turn may lead to lowered spawning rate or fertility. Spawning the females in the maturation site and transporting the eggs to the hatchery site is being considered as an alternative. Egg transport costs may be reduced to a minimum by using eggs from ablated spawners, transported at high density with no aeration. Experiments on higher egg densities as well as on transport of nauplii should, however, be undertaken.

Fish sauce from two species of unexploited deep sea fish Peristedion adeni and Peristedion weberi

Two species of unexploited deep sea fish Peristedion adeni and Peristedion weberi caught from the Exclusive Economic Zone of India were subjected to fermentation at ambient temperature (30 ± 2°C) in the presence of salt in 4:1 ratio. Periodical analysis of the fermented product was carried out up to one year. The sauces had brownish yellow colour and conformed to special grade of the standards prescribed by the Food and Drug Administration. Further solubilization of protein after 9 month's maturation was not appreciable.

Reprogramming following somatic cell nuclear transfer in primates is dependent upon nuclear remodeling

BACKGROUND: Somatic cell nuclear transfer (SCNT) requires cytoplast-mediated reprogramming of the donor nucleus. Cytoplast factors such as maturation promoting factor are implicated based on their involvement in nuclear envelope breakdown (NEBD) and prema