• Development and Reproduction
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• v.12 no.1
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• pp.77-86
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• 2008

This study was carried out to investigate the effect of leukemia inhibitory factor (LIF) on the derivation of mouse ES cells from isolated blastomeres. Two-cell stage mouse embryos were obtained from superovulated BDF1 female mice. Collected embryos were cultured to blastocyst stage in culture medium supplemented with 0, 1,000, 2,500 or 5,000 U/mL of LIF. Cultured blastocysts were examined by counting the number of cells in the inner cell mass (ICM) and trophectoderm (TE) using differential staining method. When 2-cell embryos were cultured with 2,500 U/ml of LIF, the cell numbers of ICM significantly increased in comparing with those of the control($21.0{\pm}4.0$ vs. $15.9{\pm}5.0$, P<0.01) and 1,000 U/mL of LIF-containing group ($21.0{\pm}4.0$ vs. $16.6{\pm}4.9$, P<0.05). We used an ES cell establishment medium with 20% Knockout Serum Replacement and 0.01 mg/mL ACTH instead of fetal bovine serum. Establishing efficacy of ES cell lines were the highest in 2,500 U/mL of LIF-containing group as 36.7% (11/30). This culture medium was applied to the culture of isolated blastomeres and to derivate ES cell lines. Three ES cell lines (21.4%) from isolated blastomeres of 2-cell stage embryos were established. In further experiments, we could establish one ES cell line (4.0%) from single blastomere of 4-cell stage embryo. The subcultured ES cells and their embryoid bodies were characterized by analyzing gene expression for undifferentiation and differentiation marker gene using immunocytochemistry and RT-PCR. In conclusion, LIF supplementation in culture medium could increase the cell number in ICM of blastocysts and support derivation of ES cell lines from isolated blastomeres.

• Development and Reproduction
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• v.14 no.4
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• pp.269-279
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• 2010

Some characteristics of germ cell differntiations and the function of accessory cells during spermatogenesis, and mature sperm ultrastructure in male Protothaca (N.) jedoensis were investigated by transmission electron microscope observations. The morphology of the spermatozoa of this species has a primitive type and is similar to those of other species in the subclass Heterodonta. Accessory cells, which are connected to adjacent germ cells, are involved in the supplying of the nutrients for germ cell development. The morphologies of the sperm nucleus and the acrosome of this species are the cylindrical type and cap shape, respectively. Spermatozoa are approximately $46{\sim}50{\mu}m$ in length including a long sperm nucleus (about $2.44{\mu}m$ in length), an acrosome (about $0.45{\mu}m$ in length), and tail flagellum (about $42{\sim}46{\mu}m$). The axoneme of the sperm tail shows a 9+2 structure. As some characteristics of the acrosomal vesicle structures, the basal and lateral parts of basal rings show electron opaque part (region), while the anterior apex part of the acrosomal vesicle shows electron lucent part (region). These characteristics of the acrosomal vesicle were found in the family Veneridae and other several families in the subclass Heterodonta. These common characteristics of the acrosomal vesicle in the subclass Heterodonta can be used for phylogenetic and systematic analysis as a taxonomic key or a significant tool. The number of mitochondria in the midpiece of the sperm of this species are four, as one of common characteristics appear in most species in the family Veneridae and other families in the subclass Heterodonta. However, exceptionally, only three species in Veneridae of the subclass Heterodonta contain 5 mitochondria. The number of mitochondria in the sperm midpiece can be used for the taxonomic analysis of the family or superfamily levels as a systematic key or an important tool.

• Reproductive and Developmental Biology
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• v.37 no.1
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• pp.23-27
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• 2013

This study was carried out to investigate the influence of artificial insemination (AI) failure in 1,619 Korean native cows at Gangwon East area, Korea. The average AI failure rate was 37.02% in the cows and the highest rate was 40.85% in Yangyang-city. Based on the parity in the cows, the AI failure rate was 49.14% and 29.91% in the first and fifth parity cows, respectively. Whereas cows until fifth parity were decreased in AI failure, cows with sixth or more parity showed an upturning AI failure trends with the increasing of parity number. AI failure rate incidence according to the rump fat thickness measured by ultrasound was 28.9% and 33.4% at 5 mm to 10 mm and over than 15 mm, respectively. There was a positive correlation (0.2186) between AI failure rate of mother and that of their offspring cows. That is, offspring of dams with high AI failure rate showed also higher AI failure than those of dams having lower AI failure rate. In conclusion, these results indicate that the AI failure rate was closely related to the rump fat thickness, parity number, and conception rate of mother cows. In addition, these results might strengthen the basis to improve the reproductive performance in Korean native cows.

• The Korean Journal of Ecology
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• v.24 no.3
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• pp.157-168
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• 2001

Herbicides have been used to control weeds for decades. If detoxification upon exposure to herbicides requires considerable amounts of energy, it could affect the pattern of resource allocation to growth and reproduction of crops. We examined the effects of three levels of a herbicide (Control, Low, and High) on germination, growth and reproductive characters of Glycine max treated twice, i.e., before and after seed germination. Since flowering time of G. max was separated into two groups, flowering time was also considered as a variable in this study. The rate of seed germination tended to be higher at the low level of herbicide compared to other levels. Chlorosis and shape variation of leaves were apparent after the second herbicide treatment, but completely disappeared after six weeks of treatment. The herbicide effects on growth characters were somewhat different between early and late flowering plants, but plants treated with both low and high levels of herbicide reduced their growth compared to those in the control group regardless of flowering time. Plants at the high level of herbicide exhibited the highest growth rate later in the season, suggesting that plants compensated to some extent for reduced growth. However, growth reduction among plants at the high level of herbicide was persistent until the end of growing season. Among plants flowered late in the season, plants in the control level bore a higher number of nodules per plant than those in other levels; such a pattern did not exist among plants flowered early in the season. Plants treated with low and high levels of herbicide produced a lower number of flowers than those in the control. Thus, the herbicide examined affected not only the growth and reproductive characters of non-target crops but also the development and growth of root nodules.

• Korean journal of applied entomology
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• v.37 no.2
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• pp.143-149
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• 1998

Development and life table statistics of indian meal moth (Plodia interpunctella, Hiibner)on brown rice (Sativa oryzae L., var 'Ilpum' ) were tested under five different temperatures (17, 20, 25,28 and 3 2 f 0.5"C). The developmental response of females to the temperature was not significantlydifferent from that of males. In the tested temperature range, developmental period and life span of adultmoth decreased as the temperature increased and ranged from 149.9f30.4 to 38.1 k5.6 days and from19.4f 5.1 to 6.9k2.0 days at 17$^{\circ}$C and 32"C, respectively. Emergence rate increased with the increaseof temperature and ranged from 13.0f 6.2% at 17$^{\circ}$C to 49.2f 25.9% at 32$^{\circ}$C. However, hatching ratecurve in relation to the temperature was dome shape with the peak of 73.8 k5.37~a t 25"C, suggesting thathatching is inhibited by high temperature above that temperature. As the temperature increased, femalesconcentrated their oviposition on the second day after emergence. In the temperature range of 17 SIM 25"C, the number of eggs laid per female were not related to the temperature and ranged from 133.4f 37.6to 154.3k57.4. But the number of eggs laid per female decreased at 32$^{\circ}$C which suggests closerelationship with hatching ability. The net reproduction rate was highest at 28$^{\circ}$C and followed by those at25$^{\circ}$C and 20$^{\circ}$C. However intrinsic rate of natural increase of the moth population on brown rice wasestimated to be highest at 32$^{\circ}$C (0.065 per day), probably due to the short developmental period, highemergence rate and the concentrated oviposition of females on earlier days of the emergence.ition of females on earlier days of the emergence.

• Reproductive and Developmental Biology
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• v.28 no.2
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• pp.119-126
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• 2004

The present study was carried out to examine the effect of four different media (NCSU (North Carolina State University)-23, PZM (Porcine Zygotes Medium)-3, PZM-4 and TCM (Tissue Culture Medium)-l99) and two oxygen concentrations (39 , 5% $O_2$, 5% $CO_2$ and 90% $N_2$, 5% $CO_2$ in air) on in vitro production of porcine IVM/IVF embryos. The results were summarized as follows: The rates of GVBD and nuclear maturations were not significantly different (p>0.05) for 44 hours of culture with four media in two oxygen concentrations. The rates of polyspermy, penetrated sperm(s) and male and female prouclei formation were not significantly different (p>0.05). among four media in two oxygen concentrations. The cleavage rates were not significantly different (p>0.05) among four media in two oxygen concentrations. At day 7 under gas atmosphere of 5% $O_2$, 5% $CO_2$ and 90% $N_2$, the blastocyst formation was significantly higher (p<0.05) in PZM-3 (19.9$\pm$2.4) than other media. Also, NCSU-23 medium gave high rate of blastocyst formation at day 7 under gas atmosphere of 5% $CO_2$ in air (p<0.05). Based on the result of differential staining of porcine blastocyst at dat 7, inner cell mass cell and total cell numbers were not significantly different (p>0.05) among four media in two oxygen concentrations. However, the observed total cell number was higher in PZM-3 medium (36.8$\pm$6.5) than other madia. In conclusion, these results suggested that in vitro production of porcine embryos in PZM-3 medium under a gas atmosphere of 5% $O_2$, 5% $CO_2$ and 90% $N_2$ was effective on the blastocyst formation rate and total blastocyst cell number.

• Korean journal of applied entomology
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• v.61 no.3
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• pp.423-434
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• 2022

The two previously developed artificial diets (N4 and N6) used for rearing Spodoptera frugiperda (Noctuidae) larvae, were selected as highly-fit ones for rearing Mythimna loreyi larvae. Almost all biological characteristics were not significantly different between the colonies reared on the two diets at 25℃ and 15:9 h (light:dark) photoperiod. The developmental periods were 4.9-5.2 days for eggs, and 22.3-23.2 days for larvae. The pupal period and weight were different between the sexes in each diet colony. The pupal periods in females and males showed 12.6-12.8 days and 14.1-14.5 days, respectively. The pupal weights were ca. 345 mg for females and ca. 380 mg for males. The pupation and emergence rates were ca. 91-94%, and ca. 91-95%, respectively, without significant differences between the two colonies. The pre-oviposition and oviposition periods were 3.4 days and 4.7-4.8 days, respectively. The adult longevity was 8.2 days in females and 10.3-12.4 days in males. Total offsprings produced were found to be 724-847 larvae on an average with ca. 1,400 maximum larvae. In the life table analysis, the intrinsic rates of increases (0.1181 for N4 and 0.1253 for N6) were not significantly different between the two colonies. Individual differences in the larval instar number 5 and 6 were found within a diet colony. The ratios of 5-instar larvae were ca. 22% in N4 colony and ca. 7% in N6 colony. The larval period of 6-instar larvae was longer than that of 5-instar larvae. Width of head capsule in larvae varied from ca. 309 ㎛ for 1st instar to ca. 3,065 ㎛ for 6th instar. Body lengths measured from ca. 2.0 mm for 1st instar to ca. 29.1 mm for 6th instar. Larvae of M. loreyi and M. separata were found at the same time in a maize field during June and July, 2020.

• Korean Journal of Environmental Biology
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• v.40 no.3
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• pp.281-289
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• 2022

The effect of five different constant temperatures (18, 21, 24, 27, and 30±1℃) and a photoperiod of 14 : 10 (L :D) h on the reproduction parameters of Spodoptera frugiperda was studied. The longevity of adult female S. frugiperda decreased with increasing temperature (22.4 days at 21℃ and 13.9 days at 30℃) but not at 18℃. The pre-oviposition period and oviposition period was the shortest at 30℃ compared to the other temperatures. The total fecundity egg count was 887.4, 1,246.4, 1,348.9, 1,154.9, and 1,034.2 at 18, 21, 24, 27 and 30℃, respectively, during its life span. The survival rate of female S. frugiperda decreased rapidly after 13 days at 18℃, after 14 days at 21℃, after 15 days at 27℃, and after 9 days at 24℃, and 30℃. On the third day after the start of oviposition, 50% of the total fecundity was accomplished. In corn fields at less than the 10-leaf stage, the distribution of S. frugiperda egg masses was observed in the middle and lower plant regions, corresponding to 46.8% and 41.4% of the total egg masses, respectively. Egg masses were mostly found on the underside of the leaf blade (abaxial) of corn(66.7%). After releasing S. frugiperda adults on May 12, May 17, May 25, and May 30, the number of eggs per egg mass was 89.9, 88.5, 126.6, and 127.9, respectively. Egg masses of the subsequent generations of S. frugiperda were observed from late June, and the number of eggs per egg mass was 155.8 in late June, 270.7 in early July, and 303.5 in mid-July.

• Korean Journal of Animal Reproduction
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• v.27 no.3
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• pp.215-223
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• 2003

This study was to investigate the effects of fertilization time and culture medium of pig oocytes matured in-vitro by liquid boar sperm. The sperm rich fraction (30∼60 ml) was slowly cooled to room temperature (20∼23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min 800 ${\times}$ g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of the LEN diluent to provide 1.0${\times}$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$. The medium used for oocyte maturation was TCM-199 supplemented with 26.19 mM sodium bicarbonate, 0.9 mM sodium pyruvate, 10 $\mu\textrm{g}$/ml insulin, 2 $\mu\textrm{g}$/ml vitamin B$_{12}$ , 25 mM HEPES, 10 $\mu\textrm{g}$/ml bovine apotransferrin, 150 $\mu$M cysteamine, 10 IU/ml PMSG, 10 IU/ml hCG, 10 ng/ml EGF, 0.4% BSA, 75 $\mu\textrm{g}$/ml sodium penicillin G, 50 $\mu\textrm{g}$/ml streptomycin sulfate and 10% pFF. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 1, 3, 6 and 9 h in 500 ${mu}ell$ mTBM fertilization media with 1.0${\times}$10$^{6}$ sperm/ml concentration, respectively. Thereafter, oocytes were transferred into 500 ${mu}ell$ NCSU-23, HEPES buffered NCSU-23, PZM-3 and PZM-4 culture media, respectively, for further culture of 6, 48 and 144 h. The rates of sperm penetration and male pronuclear formation were higher in the fertilization times for 6 and 9 h than in those for 1 and 3 h. The rates of cleaved oocytes were higher in the fertilization times for 6 and 9 h (85.0 and 84.6%) than in those for 1 and 3 h (61.1 and 76.8%). The percentage of blastocyst formation from the cleaved oocytes was highest in the fertilization time for 6 h (33.6%) than in that for 1, 3 and 9 h (11.4, 23.0 and 29.6%). Mean cell numbers per blastocyst were 32.9, 27.6, 26.3 and 24.4 in the fertilization times for 6, 9, 3 and 1 h, respectively. The rate of blastocyst from the cleaved oocytes and the number of cells per blastocyst were higher in HEPES buffered NCSU-23 culture medium than in NCSU-23, PZM-3 and PZM-4 culture media. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend the coincubation time of 6 h in 500 ${mu}ell$ TBM fertilization medium with 1${\times}$10$^{6}$ sperm/ml concentration and the HEPES buffered NCSU-23 culture medium for in-vitro fertilization of pig oocytes matured in-vitro.

• Korean Journal of Animal Reproduction
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• v.22 no.4
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• pp.425-435
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• 1998

To improve the efficiency of nuclear transplantation in bovine, in this study the development in vitro of nuclear transferred (NT) embryos was compared by different activation regimens of the enucleated oocytes. The effect of developmental stage and culture system of donor nuclei on fusion and development in vitro of NT embryos were also evaluated. Oocytes were collected from Hanwoo ovaries obtained from slaughterhouse and matured in Ham's F-10 supplemented with hormones. After 20~22 h maturation, the oocytes were vortexed to be free from cumulus cells and subsequently their nucleus and the first polar body were removed. Enucleated oocytes were divided into 3 groups for activation; the oocytes of group I were activated with ionomycin for 5 min and subsequently incubated in 6-dimetylarninopurine (DMAP) for 4 h, Those of group II were treated with DMAP for 4 h at 39 h after onset of in vitro maturation (IVM) and those of group III were kept in room temperature ($25^{\circ}C$) for 3 h at 39 h after onset of IVM. After in vitro fertilization (IVF) the embryos for muclear donor were cultured either by group culture (20 embryos /50 ${mu}ell$ drop) or individually (1 embryo /50 ${mu}ell$ drop) for 4 day and 5 day. At day 4 and 5 after IVF, blastomeres were separated in calcium-magnesium free medium, and then classified into small (day 5: $\leq$ 38 ${\mu}{\textrm}{m}$, day 4: $\leq$ 46 ${\mu}{\textrm}{m}$) and large (day 5 : $\geq$ 38 ${\mu}{\textrm}{m}$, day 4 ; $\geq$ 46 ${\mu}{\textrm}{m}$). The separated blastomeres were replaced into enucleated and activated recipient cytoplasm. The blastomere-oocyte complexes were fused by electrically. The NT embryos were cultured in TCM-199 containing 10% FCS in 39$^{\circ}C$, 5% $CO_2$ incubator for 7 day. The results obtained were summarized as follows; There were no differences in fusion and development to blastocyst between groups as group I (68%, 10%), group II (75%, 14%) and group III (73%, 9%), respectively. However, the cell number in blastocyst of NT embryos in group III were significantly fewer than in the other groups (P＜0.05). No differences in fusion and development to blastocyst were found between individual or group cultured and between small or large blastomeres of day 4 and day 5 donor embryos. From these results, it was concluded that the combination of ionomycin and DMAP, or treatment of DMAP at 39 h after onset of IVM were useful for the efficient of production of NT bovine embryos, and the individual cultured embryos could be simply used as donor nuclei for NT bovine embryo.