• Journal of Aquaculture
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• v.4 no.1
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• pp.31-66
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• 1991

This paper deals with the reproductive ecology e.g., number of the pre-spawning moults, morphological characteristics of the pre-spawning moult the common moult, daily ration druing a molting cycle mating behavior, structures of spermatozoa and spermatophore, structure of vas deferens, mechanisms of the oviposition and brooding into the egg-chambers, a suitable time for the artificial mating and fertilization, time sequence of the oviposition and brooding into egg-chambers from the copulation, responses to temperature and chlorinity on the egg development and hatching, effect of temperatures on duration of egg development, physical mechanism of the egg hatching, to make an attempt for the artificial spawning and brooding to establish a suitable system of the artificial seedling-production for the aquaculture. 1. Females molted commonly $8{\~}10$ times at an interval of $17{\~}18$ days at $28^{\circ}C,\;3.26\~4.35\%_{\circ}$ while the prespawning moltings were $4{\~}5$ times at an interval of $13{\~}14$ days. The suitable state for artificial copulation was within 14 hours elapsed from the prespawning moltings (most suitable state was within 8 hours). Males discharged a gelatinous spermatophore and placed it on the females sternum during copulation. Oviposition was seen $6{\~}17$ hours after copulation. External fertilization was considered to take place at oviposition. Fertilized eggs held in egg-chambers forming between pleopods were about $5000{\~}6000$ in females those sizes about 6.5 cm in body length. 2. Eggs immediately after oviposition were elliptic shape, measuring $0.58{\times}0.48$ mm up to hatching. Their sizes increased with egg development and finally reached $0.85{\times}0.54$ mm up to hatching. The relationship between the long axis of the egg(Y in U) and days elapsed(X) was expressed as Y= 5.60194 + 0.007358X. The eggs performed superficial cleavage and their cleavage furrows became visible at the 4-daughter-nucleus stage. The eggs showed normal development up to hatching at water temperature range of $22{\~}30^{\circ}C$ (optimum temperature : $26{\~}28^{\circ}C$) and at chlorinity range of $0.00\~6.64\%_{\circ}$ (optimun chlorinity : $2.21{\%}_{\circ}$). The relationship between incubation period (Y in days) and water temperature(X in $^{\circ}C$) could be expressed as Y= 50.803-1.3555X. The eggs hatched $12{\~}13$ days after oviposition at $28.0{\~}28.6^{\circ}C$ 3. The pre-spawning moltings were appreciably different in the morphologic structure from those of common moltings. Breeding setae and dresses were formed on the thoracic regions, abdominal epimerae and the bases of the first to fourth pleopods in order to prepare and support oviposition, transfering and supporting eggs in egg-chambers up to hatching. These supplementary breeding organs were observed only at reproductive seasons.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.2
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• pp.123-128
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• 1998

Implantation rates remain low following human in vitro fertilization (IVF). Suboptimal culture conditions may limit the ability of embryos to hatch as blastocysts, and artificial opening of the zona pellucida has been proposed as a means to promote subsequent hatching (assisted hatching). In this study, assisted hatching (AH) by zona drilling using acidic Tyrode's solution was performed in 320 patients, due to their age of more than 38 years (group A), the thick zona pellucida (group Z; $ZP\geq0.18{\mu}m$), and failures in implantation more than 3 times in previous IVF-ET trial (group P). This study was designed firstly, to study the effects of AH on the outcomes of IVF-ET according to the indications and secondly, to verify the appropriate application of AH. The results were as follows; 1. There was no difference in pregnancy rate between AH group (26.6%) and non-AH group (26.5%). 2. Assisted hatching (AH) showed significantly higher pregnancy rate of the patients with thick zona pellucid a than those of the patients with age factor and with the history of repeated implantation failure. But in the patients with age factor only, AH resulted in higher pregnancy rate. 3. Interestingly, the patients with complex factors including zona factor (Z: 33.9%; ZA: 30.4%; ZP: 31.6%; ZAP: 21.4%) showed higher pregnancy rates than other complex factors excluding zona factor (A: 24.4%, P: 0%; AP: 10.8%). From these results, AH is more helpful to the patients with thick zona pellucida rather than patients with older age and/or previous repeated implantation failure.

• Korean Journal of Ichthyology
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• v.36 no.1
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• pp.20-29
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• 2024

Hemitripterus villosus, a promising aquaculture fish species, is facing declining stocks. This study aims to provide normative standards for skeletal development to address persistent skeletal deformities in farmed fish. Specimens utilized in the study underwent artificial insemination with captured fish, and the resulting larvae and fry were preserved in a formalin solution. The skeletal ossification process commenced immediately after hatching, affecting the parasphenoid, premaxillary, maxillary, and dentary structures at an average total length of 13.65±0.71 mm (n=5). By sixty-five days post-hatching, ossification extended to the ethmoid and supraorbital, completing the head's development at an average total length of 21.24±0.50 mm (n=5). Clavicle ossification occurred at seven days post-hatching, corresponding to an average total length of 14.61±0.52 mm (n=5). At forty-four days post-hatching, the ossification of 4 actinosts took place, completing the shoulder girdle, with an average total length of 18.15±0.61 mm (n=5). Vertebral ossification initiated at ten days post-hatching, with an average total length of 14.80±0.65 mm (n=5). By fifty-four days post-hatching, 39 vertebral columns were ossified, reaching an average total length of 18.67±0.54 mm (n=5). Vertebral development was complete at sixty days post-hatching, with an average total length of 20.25±0.45 mm (n=5). This study sheds light on the skeletal development of H. villosus, providing valuable standards and fundamental data for understanding skeletal deformities in this species.

• International Journal of Industrial Entomology and Biomaterials
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• v.26 no.1
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• pp.61-66
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• 2013

Baculum elongatum has several peculiarities such as parthenogenesis, unique external features, changing body color, and dropping oviposition, which makes it a potential economically useful insect. Oviposition was performed by females without fertilization by the sperm of males. The oviposition period was $42.2{\pm}22.7$ days and the number of eggs per female was $109.5{\pm}70.5$ eggs. The hatch rate was 73.3% at $25^{\circ}C$ and 66.7% after low temperature treatment ($8^{\circ}C$ for 60 days). In nature, B. elongatum overwinters as an egg, but it can be assumed from the results that cold temperatures were not required for hatching. The hatch rate was 98.2% in the treatment using floral foam, fermented sawdust, and leaves. The developmental period was $100.9{\pm}4.2$ days for eggs, $55.3{\pm}4.6$ days for nymphs, and $49.7{\pm}16.0$ days for adults. The length of the eggs was $0.33{\pm}0.0$ cm and the lengths of the nymphs were clearly distinguishable according to the instar stage. Clover(Trifolium repens) was an excellent diet as it was similar to the host plant and could be used as an alternative diet. The rate of reaching adulthood for the insects was 66.7% on an artificial diet containing 25% acacia leaves. For sustainable mass rearing of nymphs or adults of B. elongatum, a natural diet could be used such as acacia (Robinia pseudoacacia), white oak (Quercus aliena), chestnut (Castanea crenata var. dulcis), and bush clover (Lespedeza bicolor), or an alternative diet such as clover or artificial diet.

• Clinical and Experimental Reproductive Medicine
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• v.38 no.2
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• pp.87-92
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• 2011

Objective: This study aimed to determine the safety and clinical effect of artificial shrinkage (AS) in terms of assisted hatching of fresh blastocysts. Also, we evaluated the correlation between patient age and the effect of AS on clinical outcome. Methods: Two AS methods, using a 29-gauge needle and laser pulse, were compared. Seventy-three blastocysts were shrunk using a 29-gauge needle and the same number of other blastocysts were shrunk by a laser pulse. We evaluated the shrunken blastocysts hourly and considered them viable if they re-expanded >70%. Blastocyst transfer cycles (n=134) were divided into two groups: a control group consisted of the cycles whose intact embryos were transferred (n=100), while the AS group consisted of the cycles whose embryos were replaced following AS (n=34). The implantation and pregnancy rates of the control group and AS group were compared ($p$ <0.05). Results: The re-expansion rates of the 29-gauge needle and laser pulse AS groups were similar (56 [76.7%] vs. 62 [84.9%], respectively). All of the remaining shrunken blastocysts were re-expanded within 2 hours. There was no degeneration of shrunken blastocysts. The total and clinical pregnancy rate of the AS group (23 [67.6%]; 20 [58.8%], respectively) was significantly higher than that of the control group (47 [47.0%]; 39 [39.0%], respectively). In the older patient group, there was no difference in the clinical outcomes between the AS and control groups. Conclusion: These results suggest that AS of blastocoele cavity, followed by the transfer, would be a useful approach to improve the clinical outcome in cycles in which fresh blastocyst stage embryos are transferred.

• Development and Reproduction
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• v.20 no.1
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• pp.31-40
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• 2016

This study was conducted in order to examine the egg development in red spotted grouper, Epinephelus akaara and the morphological development of its larvae and juveniles, and to obtain data for taxonomic research. This study was conducted in June 2013, and 50 male and female fish were used for the study. One hundred ${\mu}g/kg$ of LHRHa was injected into the body of the fish for inducing spawning, and the fish were kept in a small-sized fish holder ($2{\times}2{\times}2m$). Eggs were colorless transparent free pelagic eggs, 0.71-0.77 mm large (mean $0.74{\pm}0.02mm$, n=30), and had an oil globule. Hatching started within 27 h after fertilization. Pre-larvae that emerged just after hatching were 2.02-2.17 mm in total length (mean $2.10{\pm}0.11mm$), their mouth and anus were not opened yet, and the whole body was covered with a membrane fin. Post-larvae that emerged 15 days post hatching were 3.88-4.07 mm in total length (mean $3.98{\pm}0.13mm$), and had a ventral fin with two rays and a caudal fin with eight rays. Juveniles that were formed at 55 d post hatching, were 31.9-35.2 mm in total length (mean $33.6{\pm}2.33mm$), with red color deposited over the entire body, and black chromophores deposited in a spotted pattern. The number of fin rays, body color, and shape were the same as that in the adult fish.

• Development and Reproduction
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• v.18 no.1
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• pp.13-23
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• 2014

This study is conducted to monitor the morphological developmental features of the egg development, larvae and juvenile of Epinephelus septemfasciatus, the fertilized eggs were gotton using artificial insemination. Matured parents are collected from marine caged fish farms in Geomun-ri, Samsan-myeon, Yeosu-si, Jeollanamdo Korea in June 2012. The fertilized eggs were pelagic eggs containing one oil globule, and measured 0.81~0.89 mm ($0.85{\pm}0.04mm$, n=50) in diameter. In regard to rearing environment, the water temperature is $21.0{\sim}23.0^{\circ}C$ and the salinity is 32.0~33.2‰. Hatching was observed from 48 hours after fertilization, the mouth and anus of prelarvae was not opened but had egg yolk at newly hatched. 4 days after hatching, the mouth and anus of postlarvae was opened and began to eat Rotifer and was measured 2.40~2.49 mm ($2.45{\pm}0.03mm$ n=10) in total length. 12 days after hatching, postlarvae was measured 3.77~4.67 mm ($4.27{\pm}0.33mm$) in total length, its the second pole tide of dorsal fin and the first pole tide of pelvic fin was extended longitudinally. 71 days after hatching, juvenile was measured 40.5~45.4 mm ($42.6{\pm}2.04mm$) in total length. Seven bands were observed in body, and pole tides of dorsal and pelvic fins were shortened.

• Development and Reproduction
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• v.17 no.4
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• pp.345-351
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• 2013

This study examines the effects on fertilization rate (FR), hatching rate (HR), and normal individual rate after artificial fertilization using frozen thawed sperm according to the cryoprotectant (DMSO) concentration and the period of cryopreserved sperm of longtooth grouper, Epinephelus bruneus. Performing artificial fertilization using frozen-thawed sperm, after freezing the sperm at different DMSO concentration of 5.0%, 7.5%, 10.0% respectively, FR were (DMSO 5.0%: $99.5{\pm}0.8%$, DMSO 7.5%: $99.5{\pm}0.7%$, and DMSO 10.0%: $99.6{\pm}0.6%$). The results are not significantly different from the control fresh sperm (100%). HR also (DMSO 5.0%: $96.2{\pm}2.3%$, DMSO 7.5%: $95.3{\pm}3.6%$, 10.0%: $96.6{\pm}1.8%$) were not significantly different in each group. The normal individual rate after hatching using with control fresh sperm ($98.4%{\pm}0.5$) and DMSO concentration level of 5.0% ($97.8{\pm}0.1%$) were not significantly different. However, with 7.5% ($97.2{\pm}0.6%$) and 10.0% DMSO concentrations ($95.9{\pm}0.2%$) are lower than the normal individual rate after hatching observed in the control and 5.0% DMSO. Performing artificial fertilization using frozen-thawed sperm at different frozen period (2 days, 2 years, and 3 years), 10% DMSO FR and HR of 3 years (FR; $66.8{\pm}1.8%$, HR: $82.0{\pm}12.9%$) and 2 years (FR; $78.5{\pm}14.8%$, HR: $79.3{\pm}0.6%$) cryopreserved sperm were lower than control (FR; 100%, HR: $91.1{\pm}3.6%$) and 2 days cryopreserved sperm (FR; $99.6{\pm}0.6%$, HR: $96.6{\pm}1.8%$). These results suggest suitable DMSO concentration ranges of cryopreservation sperm for E. bruneus is 5 to 10% and with 2 to 3 years cryopreservation period, cryopreservation sperm can be useful for seed production.

• Journal of Embryo Transfer
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• v.25 no.1
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• pp.9-14
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• 2010

This study was conducted to find out the effects of artificial shrinkage (AS) on post-thaw development of bovine embryos. The blastocoelic cavity of blastocyst was punctured to remove its fluid contents and then incubated in the holding medium (HM) for 10 min. The punctured and non-punctured (control) blastocysts were equilibrated in vitrification solution 1 (VS1; TCM-199+20% FBS+10% EG) for 5 min and vitrification solution 2 (VS2; TCM199+20% FBS+35% EG+5% PVP+0.5 M Sucrose) for 1 min and vitrified by direct dropping into the liquid nitrogen. Vitrified blastocysts (punctured and control) were thawed and cultured in vitro (12 hr) for studying survival and hatching rates. The levels of shrinkage were measured by the volume of the blastocyst during equilibration in VS1 (at 1, 3 and 5 min of equilibration) and VS2 (at 30 and 60 sec of equilibration) that was considering the volume of non-punctured blastocyst in HM as 100%. The levels of shrinkage were higher in punctured group (62.4, 64.6, 64.3% at 1, 3 and 5 min in VS1; 50.6 and 52.7% at 30 and 60 sec in VS2) than control group (84.8, 86.6, 86.4% at 1, 3 and 5 min in VS1; 72.1 and 68.8% at 30 and 60 sec in VS2), but within each group the levels of shrinkage were similar. The survival (90.9%) and hatching (50.0%) rates of vitrified blastocysts at 12 hr post-thaw were higher in punctured group than that in control group (76.9% and 0.0% respectively). We confirmed that vitrification solutions (VS1 and VS2) have no toxic effect on the survival of blastocysts because the survival rates of blastocysts exposed to VS1 and VS2 for 24 hr were similar between punctured and control groups (94.3 vs. 96.0%; p>0.05). In conclusion, the preliminary data show that AS of blastocyst may improve survival and hatching rate after thawing.

• Korean Journal of Ichthyology
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• v.28 no.1
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• pp.9-18
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• 2016

Histological and morphological development of the digestive tract of sevenband grouper were observed from after hatching to 60 days. Fishes were fed with rotifer (Brachionus rotundiformis) and chlorella (Chlorella ellipsoidea) after hatching from 2 to 20 days rotifer and brine shrimp (Artemia salina) in after 20 days rotifer, brine shrimp and semi-dry artificial diet in after 23 days. Histological and morphological development of ten larvae was observed by paraffin embedding method after fixing in 10% neutral buffered formalin. Sevenband grouper RLG showed characteristics of carnivorous fish by average 0.87. Larvae after hatching can't open the mouth and anus digestive tract was observed in a straight line following yolk sac. Larvae was observed feeding activity by opening the mouth and anus. Metamorphosis started 8 days after hatching. Esophagus divided four layer, and goblet cell was observed in esophagus, mid intestine and rectum. Larvae started cannibalism and it was caused by difference of growth. The inside of stomach was differentiated to cardiac orifice, body of stomach, pyloric stomach, and pyloric caeca. Goblet cell was observed all intestine. Gastric gland differentiated after hatching 28 days in stomach. Secretion of gastric juice was found at stomach and mucosal fold pyloric caeca. Even thought the inside of stomach expended and the number of gastric gland increased consistently and goblet cell in intestine and mucosa became longer, histochemical changes follow couldn't be found during transforming juveniles 38 days after hatching.