• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.103-103
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• 2002

This study was conducted to investigate cryopreservation of pearl oyster, Pinctada fucata martensii larvae. Four cooling rates (-0.25, -0.5, -0.75 and -1.0$^{\circ}C$/min.) were used to examine a proper cooling rate during cryopreservation of trochophores before seeding temperature (-12$^{\circ}C$). Seven developmental stages (early and late trochophores, early and late D-shaped larvae and early, middle and late umbo stage larvae) and different sugars (fructose, glucose and sucrose) were used to investigate optimal larval stage and effective sugar in cryopreservation of larvae. The survival rates of frozen-thawed trochophores increased at cooling rate of -1.0$^{\circ}C$/min. As larval developing, survival rate of frozen-thawed larvae increased, except umbo stage larvae, and especially late D-shaped larvae highly survived as 91%. Addition of sugar revealed positive effect on cryopreservation in this experiment and 0.2 M glucose and sucrose mixed with 2.0 M dimethyl sulfoxide significantly enhanced survival rate of larvae (P<0.05). The results of our study indicate that desirable cooling rate, developmental stages of larvae and effective sugar far cryopreservation of pearl oyster, P. fucata martensii larvae are -1$^{\circ}C$/min, late D-shaped larvae and 0.2 M glucose and sucrose, respectively.

• Korean Journal of Veterinary Research
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• v.38 no.1
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• pp.107-117
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• 1998

The cultivation for development of Ascaris suum from the second-stage larvae($L_2$) embryonated egg and the third-stage of rat-derived larvae($L_3$) recovered from lung of rats were performed to use the screening test of anthelmintics in vitro. The preparations of larvae for cultivation were that the artificially-hatched $L_2$ incubated the embryonated eggs of Ascaris suum in 0.1% formalin solution at $25^{\circ}C$ for 28 days and the rat-derived larvae($L_3$) recovered from the lung of rat infected with the embryonated eggs of Ascaris suum on 7 days after infection(DAI). The cultivation for development of Ascaris suum from the embryonated eggs($L_2$) and the rat-derived larvae($L_3$) for 14 days in RPMI medium 1640(with 5% bovine calf serum) were as follows : 1. The sizes of the liberated larvae($L_2$) which were artificially hatched from embryonated eggs with glass beads(diameter 5mm) were $190{\sim}250{\mu}m$ on 1 days in culture(DIC). The second-stage larvae were molted into third-stage larvae(early $L_3$; $250{\sim}300{\mu}m$) and the features of these larvae were first observed such as cephalic cuticle, esophageal lumen and anus etc. on 5 DIC and the sizes of late third-stage larvae were $250{\sim}450{\mu}m$ on 10 DIC. The sizes of early fourth-stage larvae($L_4$) were $500{\sim}700{\mu}m$ and the features of these larvae were more pronounced in internal organs on 15 DIC. 2. The sizes of third-stage larvae($L_3$) recovered from the lung of rats were $1,340{\sim}1,370{\mu}m$ and the feartures of cephalic cuticle, esophageal lumen, intestine, rectum, anus were visualized by inverted microscope on 1 DIC. The fourth-stage larva($L_4$) completed by third ecdysis were recognizable and sizes of early fourth-stage larvae were developed as $1,400{\sim}2,200{\mu}m$ on 5 DIC. The sizes of middle fourth-stage of larva were $1,900{\sim}2,300{\mu}m$ and the thickened epithelial rectum was observed on 10 DIC. The rectum and anus of late fourth-stage larva($L_4$ $2,500{\sim}3,200{\mu}m$) had developed completely in RPMI medium 1640 on 15 DIC.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.3
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• pp.223-231
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• 2020

Karyotype analysis is a major work in the process of triploid abalone production for the purpose of productivity and quality improvement. However, the metaphase spreads for karyotype analysis have been prepared just from the larvae at trochophore stage, which has restricted the spectrum of sample correction inhibiting more efficient analysis. Here, we investigated the feasibility of preparing metaphase spreads from the larvae at veliger stage that is the next developmental stage of trochophore. For this, diploid and triploid larvae at trochophore and veliger stages from Pacific abalone (Haliotis discus hannai) were subjected to metaphase spread preparation and its efficiencies were measured and compared each other. As the results, although the efficiencies of metaphase spread preparation were significantly lower in the larvae at veliger stage compared to the ones at trochophore stage regardless of ploidy status, we found that the preparation of metaphase spreads, which showed the clear chromosomal images containing the normal number of chromosomes, was possible from the veliger stage larvae. On the other hands, all larvae used in this study regardless of developmental stage and ploidy did not show colchicine sensitivity. Moreover, no significant difference was observed in cell cycle distribution of the cells comprising larvae between two developmental stages regardless of ploidy status. These suggested that the details of protocol to prepare metaphase spreads from abalone larvae should be optimized depending on its developmental stages. Taken together, we demonstrated the feasibility of preparing metaphase spreads from H. discus hannai veliger stage larvae for karyotype analysis.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.104-104
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• 2002

This study was performed to find out the optimal larval stage among trochophore, D-shaped and umbo stage larvae and the desirable protective additive such as fructose, glucose, sucrose and trehalose with cryoprotectant for cryopreservation of surf clam, Spisula sachalinensis larvae. Dimethyl sulfoxide and ethylene glycol were used as cryoprotectant and each cryoprotectant was made to 2.0 M with previous protective additives. The larvae were immersed in the preparations waited for 15 minutes to reach equilibration, and then frozen in a program freezer (-35$^{\circ}C$) and liquid nitrogen (-196$^{\circ}C$). The freezing rate of 1.0$^{\circ}C$ /min. was used for cryopreservation of trochophores before seeding temperature (-12$^{\circ}C$). The survival rate of frozen-thawed larvae increased as larval developing and that of umbo stage larvae was the highest as 96.1 ${\pm}$ 1.0%. The presence of lower concentration of disaccharides as sucrose or trehalose significantly enhanced survival rate when mixed with cryoprotectants (P<0.05). The results of our study indicate that desirable developmental stages of larvae and protective additive for cryopreservation are the umbo stage larvae and 0.2 M sucrose, respectively.

• Parasites, Hosts and Diseases
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• v.35 no.3
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• pp.211-214
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• 1997

Susceptibility of some vertebrates was examined to the early third-stage larvae (EL3) of Gnathostomn hispidum. The larvae collected from the Chinese loaches were infected to 4 silk carps, 3 snake heads, 3 bullfrogs. 5 mice and 9 albino rats. No worms were detected in fish. silk carps and snake heads. In 3 bullfrogs fed 30 larvae, a total of 9 EL3 was recovered in the gastrointestinal tract (8 larvae) and liver (ones). In 5 mice inferred with 50 larvae, a tolal of 37 (74.0%) advanced third-stage larvae (AdL3) was recovered from the muscle (31 larvae), liver (5 larvae) and kidney at 4 weeks after infection. In 9 albino rats iilfected with 115 larvae, a total of 40 (34.8%) AdLa was found in the muscle. The Inammalian hosts were found susceptible to the EL3 of G. hispinum from Chinese loaches.

• The Korean Journal of Ecology
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• v.21 no.6
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• pp.799-804
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• 1998

laboratory experiments were conducted to determine the effects of Dimilin, an insect growth regulator which acts to inhibit chitin synthesis, during the larval development of liocarcinus corrugatus Pennant. The larvae was exposed to control (10 ppb acetone sea water and untreated sea water solution) and five concentrations 0.1, 1.0, 5.0, 10.0 and 25.0 ppb of both TG and WP-25 formulations of Dimilin from the hatching to the megalopal stage, and the effect of Dimilin on development of the larvae were determined. Two formulations (TG and WP-25) had different effect on the different stages in L. corrugatus. and early stage larvae of L. corrugatus were more sensitive to TG than to WP-25. Concentrations of diflubenzuron >5.0 ppb are lethal to L. corrugatus larvae upon chronic exposure. Lethal concentrations are defined here as those in which less than 10% of the larvae survived to the megalopal stage. However, Dimilin (TG and WP-25) showed no significant effects on developmental time of L. corrugatus larvae.

• Parasites, Hosts and Diseases
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• v.49 no.1
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• pp.51-57
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• 2011

Four cases of gastric or intestinal myiasis are reported. The cases contain 2 males (1 child 10 years old, and 1 adult 40 years old) and 2 females (1 girl 18 years old, and 1 adult 50 years old) from Minia Governorate, Southern Egypt. Three of them, including cases no. 1, 3, and 4, were gastric myiasis, and complained of offensive hematemesis of bright red blood. Minute moving worms, larvae of the fly, were found in the vomitus. On the other hand, case no. 2 had intestinal myiasis, and complained of abdominal distention, nausea, vomiting, and diarrhea. The stool of case 2 was mixed with blood, and minute moving worms were observed in the stool. Endoscopy was performed to explore any pathological changes in the stomach of the patients. The larvae were collected and studied macroscopically, microscopically, and using a scanning electron microscope (SEM) to identify their species. Three different types of larvae were identified. The larvae isolated from case 1 were diagnosed as the second stage larvae of Sarcophaga species, and the larvae isolated from case 2 were the third stage larvae of Sarcophaga species. On the other hand, the larvae isolated from cases 3 and 4 were diagnosed as the third stage larvae of Oestrus species.

• Korean Journal of Veterinary Research
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• v.15 no.1
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• pp.51-55
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• 1975

In this study two sets of experiments were undertaken. Firstly, the embryonated eggs and first-stage larvae, sprayed in disinfected soil, were exposed to different temperatures. Then, the viabilities of them were weekly determined by inoculating the soil to earthworms, Eisenia foetida. Secondly, the infective larvae, seperated from the earthworms and suspended in tap water, were also exposed to different temperatures and their viabilities were checked microscopically at weekly intervals. The results were summerized as follows: 1. The maximum longevities of embryonated eggs and first-stage larvae were determined as 1 week at $35^{\circ}C$, over 36 weeks at $25^{\circ}C$, $15^{\circ}C$, and $5^{\circ}C$, 32 weeks at $-5^{\circ}C$, and under 1 week at $-15^{\circ}C$. 2. The mean numbers of infective larvae detected from the test earthworms were greatest at $5^{\circ}C$, and decreased with rise or fall of the temperature. 3. Infective larvae freed from the intermediate host were able to survive for 2 weeks at $25^{\circ}C$ and 3 weeks at $15^{\circ}C$. However, they lost their viabilities in a week at $35^{\circ}C$, $5^{\circ}C$, $-5^{\circ}C$, and $-15^{\circ}C$. 4. The number of living infective larvae at $15^{\circ}C$ was greater than at $25^{\circ}C$.

• The Korean Journal of Malacology
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• v.15 no.2
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• pp.115-119
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• 1999

We investigated the processes of egg and larval developments for aquaculture technique development of seedling production fo the flat oyster, Ostrea denselamellosa. Teo flat oyster of larviparous type was different from the pacific oyser (ovivarous type) because their larvae (trochophore and prodissoconch larvae) in the gill released into the seawater. The process of egg development was observed by artificial fertilization at $25^{\circ}C$, using a dissecting method. The sizes of Unfertilized eggs ranged from 80 to 90 $\mu\textrm{m}$ and fertilized eggs with globule-shape was 90-100 $\mu\textrm{m}$. The Polar body appeared after fertilization and egg cleavage began within 1 hour, reaching the blastula stage after 10 hours. The trochophore in the gill appeared 2-3 days after fertilization and grew to the prodissoconch larvae (130 140 $\mu\textrm{m}$) having a complete shell after 1-2 days. The shell of prodissoconch larvae grew to 205 220 $\mu\textrm{m}$ after 10 hours, and then they became umbo stage larvae showing oval in shape. The velum of umbo stage larvae was degenerated about 17-20 days after fertilization and grew into a pediveliger with a developed foot, at this time, the shell length size was 320 360 $\mu\textrm{m}$.

• Animal cells and systems
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• v.12 no.4
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• pp.287-295
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• 2008

Variations of larval abundance and hatch timing of Dungeness crabs, Cancer magister Dana 1852, were investigated. Dungeness crab larvae were monthly collected at 16 stations arrayed in four transects, Upper Chatham, Icy Strait, Cross Sound, and Icy Point, in southeastern Alaska from May to September 1997-2004. Larval abundance at all transects was the highest in June except in the Icy Point transect. Larval abundance was the highest in the Icy Strait transect, moderate in the Upper Chatham and Cross Sound transects, and the lowest in the Icy Point transect. Zoeae I(ZI) was predominated in May; thereafter ZI decreased and late zoeal stages occurred. In May and June, small numbers of late stage larvae unusually co-occurred with ZI in three transects. These late stage larvae may have been transported from where hatching occurs earlier. The timing of ZI occurrence varied interannually and was related to degreedays during the egg incubation period of Dungeness crabs: later larval hatching in 1997 and 2002 when temperatures were colder, while earlier larval hatching in 1998 when temperatures were warmer. The distribution patterns of Dungeness crab larvae in southeastern Alaska were markedly different from those reported from other areas of the species distribution ranges: larvae occurring much later in the year, and late stage larvae occurring in inland waters.