• Korean Journal of Animal Reproduction
• /
• v.13 no.3
• /
• pp.188-194
• /
• 1989

These experiments were carried out to investigate fertilizable and developmental ability after zona drilling the unfertilized eggs and the eggs not fertilized by the 1st insemination. The results of in vitro fertilization of the mouse eggs treated by using micromanipulation and acid tyrode's solution with capacitated epididymal spermatozoa were as follows. In the case of ovulated unfertilized eggs, according to sperm count(106, 105, 104 and 103/ml) the rates of in vitro fertlilization treated by zona drilling were 86.0%, 82.0%, 70.0% and 54.0%, respectively, and those of control were 58.0%, 52.0%, 12.0% and 8.0%, respectively. The rates of in vitro fertilization of zona drilled eggs were significantly high compared with those of control, and there were no significant difference between two groups. According to the sperm count the zona drilled eggs developed to the blastocysts were 51.4%, 40.5%, 23.3% and 17.4% and those of control were 35.7%, 26.3%, 0% and 0%, respectively. Also, in the eggs not fertilized by 1st insemination, the fertilization rates of oocytes reinseminated after zona drilling was significantly higher(83.5%) than that of control(34.7%), and the rates of polyspermy were similar. The rates of development to the blastocysts was 18.6% in the zona drilling treated eggs, and that of control was 27.3%, there was no significant difference between two groups. These results indicated that oocytes not fertilized by 1st insemination as well as ovulated unfertilized eggs could be fertilized, improve fertilizing rates by zona drilling treatment, and development potential were normal.

• Applied Microscopy
• /
• v.51
• /
• pp.3.1-3.6
• /
• 2021

We examined the morphology of fertilized egg and ultrastructures of fertilized egg envelopes of dwarf rainbowfish (Melanotaenia praecox) belong to Melanotaeniidae using light and electron microscopes. The fertilized eggs were spherical with adhesive filament, transparent, demersal, and had a narrow perivitelline space and small oil droplets. The size of fertilized egg was 1.02 ± 0.18 mm (n = 30), and there were two kinds of adhesive filament on the fertilized eggs. The long and thick (diameter 12.22 ± 0.52 ㎛, n = 20) adhesive filaments were only at the area of animal pole, and short and thin (diameter 1.99 ± 0.23 ㎛, n = 20) adhesive filaments were around the long filaments. A micropyle was conical shaped with adhesive filament and located near the animal pole of egg. The outer surface of fertilized egg was rough side. Also, the total thickness of the fertilized egg envelope was about 7.46 ± 0.41 ㎛ (n = 20), the fertilized egg envelope consisted of two layers, an inner lamellae layer and an outer layer with high electron-density. And the inner layer was 8 layers. Collectively, these morphological characteristics and adhesive property of fertilized egg with adhesive filaments, and ultrastructures of micropyle, outer surface, and section of fertilized egg envelope are showed species specificity.

• Applied Microscopy
• /
• v.43 no.1
• /
• pp.14-20
• /
• 2013

The leopard danio, Danio rerio var. frankei is a spotted color morph of the zebrafish, Danio rerio caused by a pigment mutation. The structural differences of fertilized egg and egg envelope are poorly documented. To clarify this, we compared the fertilized egg morphology and ultrastructures of surface structures, the micropyle and the cross section of fertilized egg envelopes of zebrafish and leopard danio, variation species of zebrafish using a light and electron microscopes. Although the fertilized egg sizes were different, the external shapes of the fertilized eggs of two species couldn't be differentiated under the light microscope. The characteristics of fertilized eggs, such as a spherical shape, a non-adhesive quality and a large perivitelline space, were shown to be related to spawning habit. In ultrastructure of fertilized egg envelope, there is no morphological difference of micropyle between two species. By contrast, the ultrastructure and the numbers of knob-like structures and semihemisphere-like structures per unit area on the outer surface, and the number of lamellae of inner layer on the fertilized egg envelope section displayed definite species specificity. Collectively, our data indicate that the ultrastructure of fertilized egg envelope in the zebrafish could be differentiated by species variation.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.53 no.4
• /
• pp.479-485
• /
• 2020

We evaluated the hatchability of fertilized eggs from six hybrid combinations of highly valued grouper species inhabiting temperate and warm waters, with the goal of establishing a novel hybrid with enhanced growth and viability during the culturing period in the temperate waters of Korea. Hybrid combinations with red-spotted grouper females exhibited high hatchability with high a fertilization and hatching rate of fertilized eggs and a low deformity rate of hatched larvae. Conversely, hybrid combinations with kelp grouper females had very low hatching rates and very high deformity rates; commercial production of seed from such crosses would be difficult without improving hatchability. The hatchabilities of convict grouper ♀×giant grouper ♂ and kelp grouper ♀×red-spotted grouper ♂ were lower than those of maternal purebreds, but these two hybrid combinations were expected to produce potentially large quantities of hatched larvae. In the above evaluation, promising hybrid combinations were identified for commercial production of seed. For these hybrids to contribute to the development of Korea's mariculture industry, mass production of fertilized eggs and seeds is necessary, along with the development of advanced rearing techniques, such as the identification of a suitable rearing temperature.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.54 no.1
• /
• pp.31-37
• /
• 2021

The mass production of fertilized eggs of the convict grouper Hyporthodus septemfasciatus was studied from 2013 to 2020 for industrial aquaculture. The experiment was divided into two groups. Group 1 broodstock was raised from wild-caught fry and used from 2013 to 2020. Group 2 broodstock was raised from artificially propagated fry and used from 2019 to 2020. Males used to collect sperm for artificial insemination weighed more than 7 kg. The effects of various hormones on artificial ovulation were investigated from 2013 onward. Among these, luteinizing hormone-releasing hormone analogue (LHRHa) at 100 ㎍/kg body weight showed the most effective results and was used for artificial egg collection from 2014 onward. In Group 1, the average total egg production per year, average egg production per individual, fertilization rate, and hatching rate were 26,143 mL, 609.7 mL, 93.3%, and 91.8%, respectively, and in Group 2, were 2,750 mL, 316.5 mL, 92.1%, and 90.4%, respectively. Based on these results, we showed that a large number of fertilized eggs for artificial seeding could be produced consistently. Moreover, the mass production of fertilized eggs in Group 2 establishes a foundation for the complete aquaculture cycle of H. septemfasciatus.

• Development and Reproduction
• /
• v.22 no.3
• /
• pp.289-295
• /
• 2018

Large quantity of eggs fail to be fertilized and many of fertilized eggs are unable to hatch in the eel, Anguilla japonica. Larvae of eel absorb egg yolk up to 8 days after hatching but the majority of hatched larvae die before they reach the stage of first feeding in this species. Genes of key enzymes for yolk processing (cathepsin B, D, L and lipoprotein lipase - abbreviated as ctsb, ctsd, ctsl and lpl, respectively) could be associated with egg quality. In this study, we investigated differences in the expression of these genes between floating eggs and sinking eggs, and also the relationship between the gene expressions of the enzymes and fertilization rates in the fertilized eggs obtained from artificially matured female eels. Expressions of yolk processing enzyme genes did not show significant difference between floating and sinking egg groups. Expression of ctsb decreased when fertilization rate was high. Expression of ctsd, ctsl and lpl, however, did not show any significant differences. These results suggest that ctsb expression could be an indicator of egg quality, and that some proteins prone to be digested by ctsb could be very important in the process of fertilization and normal cleavage in this species. Further study should identify these critical proteins to improve our understanding on the quality of fish eggs.

• Food Science of Animal Resources
• /
• v.37 no.5
• /
• pp.764-772
• /
• 2017

Fertilized hen eggs are rich in a variety of bioactive ingredients. In this study, we aimed to obtain an antioxidant protein from fertilized eggs and the radical scavenging abilities on 1, 1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical (${\bullet}OH$), superoxide anion ($O^{2-}{\bullet}$) were used to evaluate the antioxidant activity of the purified protein. During 20 d of incubation, the radical scavenging ability of protein extracted from fertilized eggs exhibited significantly differences and the protein on day 16 showed higher antioxidant capacity. Based on this, the antioxidant protein of the samples on day 16 were isolated for the follow-up study. With a molecular weight 43.22 kDa, the antioxidant protein was purified by Diethylaminoethyl cellulose -52 (DEAE-52) column and Sephadex G-100. The LC-MS analysis showed that the purified protein molecular weight was 43.22 kDa, named D2-S. The sequence of amino acids was highly similar to ovalbumin and the coverage reached to 84%. The purified protein showed a radical scavenging rate of $52.34{\pm}3.27%$ on DPPH and $63.49{\pm}0.25%$ on ${\bullet}OH$, respectively. Furthermore, the C-terminal amino acid sequence was NAVLFFGRCVSP, which was consistent with the sequence of ovabumin. These results here indicated that purified protein may be a potential resource as a natural antioxidant.

• Korean Journal of Ichthyology
• /
• v.22 no.3
• /
• pp.195-200
• /
• 2010

A large volume of hatching jar was tested whether it is an effective to hatch fertilized eggs of Pacific cod. The volume of hatching jar did not show any significant differences in survival of the fertilized eggs; 48.3% for the jar of 15 L and 50.4% for that of 42 L. Survival rate of the fertilized eggs in a large volume of hatching jar ranged from 40.0 to 71.2%, which was higher than those of conventional small circular tank. A higher survival could be achieved even though stocking density of fertilized eggs was as high as 5,000 mL/jar. Consequently, the tested hatching jar allowed for incubation of a greater number of eggs with higher survival rate in a much smaller space. In addition, it reduces costs and manpower, and requires a relatively small amount of water per individual unit (6,700 mL/min), and provides a way to incubate multiple rearing groups in a quarantine environment.

• Korean Journal of Animal Reproduction
• /
• v.8 no.1
• /
• pp.16-21
• /
• 1984

This experiment was carried out to compare with the nonsurgical and surgical recovery of fertilized eggs in super-ovulated rabbits. Sixty-four eggs recovered were transferred to twelve synchronized, pseudopregnant rabbits to test the viability of the eggs by surgical transfer. Each group(I, II, III) received a single subcutaneous injection of 5mg PGF2${\alpha}$/kg B.W. at 24(Group I), 48(Group II) and 72 hours (Group III) after mating, respectivity. After the administration of PGF2${\alpha}$, vaginal washings were conducted at 3, 6, 9, 12 and 24 hrs, and frequency of vaginal washing was 5 times for the each group (I, II, III). In Group (IV, V, Ⅵ), the rabbits were killed to recover the fertilized eggs from the genital tract at 24(Group Ⅵ), 48(Group V) and 72 hours (Group Ⅵ) after mating, respectively. The results obtained were as follows: 1. Of the total eggs, 69.3%, 73.4% and 66.9% were recovered for Group I, II and III, respectively from the vagina within 6 hrs after PGF2${\alpha}$ injection and particularly for Group III. 2. The rates of egg recovery versus the number of corpora lutea were 55 (51.6-60%), 35.8 (24-52.6%), 33.4 (25-47%) and 72 (70.7-73.0)%, 60.3 (50-71.4)%, 449(44.4-45.5)% in Group I, II, III and Group IV, V, Ⅵ, respectively. 3. Most of eggs recovered were one-cell stage in Group Iand Group IV. More than one half of the eggs recovered in Group II and V were over eight-cell stage, and most of the eggs were so in Group III and Ⅵ. 4. When sixty-four eggs recovered between 24 to 72 hours after mating were transferred to pseudopregnant rabbits. Three recipients were pregnant, and the rate of pregnancy was 25%.

• Applied Microscopy
• /
• v.47 no.2
• /
• pp.70-74
• /
• 2017

In the case of genus Nothobranchius, Nothobranchiidae, the morphology of fertilized eggs and ultrastructures of fertilized egg envelopes have been reported in only two fishes. Therefore it is hard for sure to these morphological characteristics show genus specificity because of lower research samples. So, we studied the morphology of fertilized egg, and compared the ultrastructures of outer surface, micropyle, and section of fertilized egg envelopes under the light and electron microscopes from the other two species, Nothobranchius foerschi and Nothobranchius rachovii, Nothobranchiidae to find out whether these structures have the species specificity or not. Both fertilized eggs were spherical, demersal and adhesive, and have a large oil droplet. The adhesive whip-like structures were distributed on the outer surface of egg envelope, and a micropyle located on the animal pole. The fertilized egg envelope consisted of two distinct layers: an outer electron-dense layer with adhesive structures and an inner lamellae layer in both species. The external shapes of fertilized egg and ultrastuctures of outer surface, micropyle, and section of fertilized egg envelope have same structure including results before. Our data indicate that these morphological characteristics of fertilized egg and fertilized egg envelope show genus Nothobranchius specificity.