• Journal of Embryo Transfer
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• v.21 no.2
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• pp.147-156
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• 2006

The objective of this study was to supply excellent genetic resources to livestock farms by transferring embryos produced by genetically superior Korean cows (Hanwoo). Eighty Hanwoo donors were superovulated with gonadotropin ($Folltrpin^(R)\;or\;Antorin^(R)$) for 4 days combined with or without progesterone releasing intravaginal device (CIDR) insertion. The collected fresh or frozen-thawed embryos were transferred to 226 farm recipients. In this study, the effect of CIDR insertion in combination with gonadotropin ($Folltrpin^(R)$) treatments initiated at the random stage of estrous cycle on embryo production was evaluated and compared to conventional superovulation protocol. Moreover, the effect of gonadotropin ($Antorin^(R)$) dose in CIDR-treated Hanwoo donors on the embryo yield was determined. In addition, the effects of embryos (fresh vs. frozen-thawed), embryo transfer person, seasons and farms on the pregnancy rate were evaluated. In Hanwoo donors, CIDR insertion in combination with $Folltrpin^(R)$ treatments regardless of estrous detection resulted in increased numbers of total ova (6.5 vs. 5.8) and transferable embryos (3.9 vs. 3.2) compared to the conventional superovulation protocol (p<0.01). In CIDR-treated Hanwoo donors, the higher dose of $Antorin^(R)$ (36 vs. 28 mg) resulted in the increased number of transferable embryos (8.3 vs. 5.4, p<0.05). The embryos (fresh 43.9% vs. frozen-thawed 23.1%) and embryo transfer person (53.9 vs. $0{\sim}16.7%$) significantly affected the pregnancy rate after embryo transfer (p<0.01). These results suggest that CIDR-based superovulation protocol may be effectively used for production of superior Hanwoo embryos and, multiple ovulation and embryo transfer in Hanwoo might be effectively applied for livestock improvement if pregmancy rate with frozen-thawed embryos and embryo transfer skill would be improved.

• Journal of Embryo Transfer
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• v.21 no.3
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• pp.191-198
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• 2006

The objectives of this study was to produce valuable offsprings of Hanwoo and Jeju black cattle using in vivo embryo production and embryo transfer techniques during 5 years ($2001{\sim}2005$) in Jeju. Two hundred and eighty six Hanwoo and sixty nine Jeju black cattles, at random stages of the estrous cycle, received a CIDR. Seven days later, the animals were superovulated with a total of 400 mg pFSH ($Folltropin^{(R)}-V$) administered twice daily in constant doses (each 50 mg) over 4 days. On the 6th administration of FSH, CIDR was withdrawn and 25 mg $PGF_{2{\alpha}}$ was administered. Cows were artificially inseminated thrice after estrous detection at 12 hr intervals. The cows received $250{\mu}g$ GnRH at the time of 2nd insemination. Embryos were recovered 7 or 8 days after the 1st insemination. Recipients were synchronized with donors by insertion of a CIDR for 7 days and administration of 25 mg $PGF_{2{\alpha}}$ at the time of CIDR removal. The collected embryos were transferred to 1,219 recipients by 6 transfer persons. The mean numbers of total ova and transferable embryos from Hanwoo and Jeju black cattle donors were 7.4 and 4.7, respectively The number of transferable embryos differed between Hanwoo (5.0) and Jeju black cattle (3.5, p<0.05), while that of total ova did not differ. Repeated superovulation treatments decreased (p<0.05) the ratio of numbers of the flushed animals vs. superovulated animals in Jeju black cattle, and the numbers of total ova and transferable embryos in Hanwoo. More transferable embryos were collected at summer (5.6) than winter (2.9, p<0.01). The mean pregnancy rate was 40%. The pregnancy.ate was affected by transfer year (2001<2004, p<0.05) and transfer person ($33.0{\sim}41.9%$, p<0.01), while not by donor (embryo) breed. These results showed that in vivo embryo preduction and embryo transfer techniques using CIDR for Hanwoo and Jeju black cattle donors as well as recipient, regardless of their estrous cycle, may enable a stable embryo production and recipient preparation.

• Journal of Aquaculture
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• v.6 no.4
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• pp.255-271
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• 1993

For the development of seedling production techniques of the freshwater crab Eriocheir japonicus, the effects of salinity and temperature on the growth of larvae of the crab were studied. Embryos hatched out as zoea larvae were measured 0.421 mm in average carapace length. Five zoea stages needed 16-26 days for metamorphosis from zoea to megalopa at $22^{\circ}C$ and $24.5\%o$. The average carapace length of the 5th zoea was 1.16 mm and that of the megalopa larvae was 1.89 mm. Each zoea stage could be identified based on both the number of plumosed seatae on the exopodite of maxilliped, and the number of spines on the posteroinner margin of tel son and also based on the rudimentary pleopods appearence. Zoea larvae fed rotifers and Artemia nauplii were healthy and metamorphosing rate from zoea to megalopa was $80\~90\%\;at\;22\~26^{\circ}C\;and\;17.5\~31.5\%o$. The relationship between larval period (Y in days) and water temperature $(X\;in\;^{\circ}C)$ is expressed as Log Y = 3.8604-1.91735 LogX. Water temperature and salinity ranges for better survival and metamorphosis of the larvae were $ 22^{\circ}C\~26^{\circ}C\;(optimum\;at\; 26^{\circ}C)\;and\;17.5\%o\~31.5\%o\;(optimum\;at\;24.5\%)$, respectively. The duration of larval stages tend to longer as salinity levels deviated from optimum particulaly at lower end. All zoea larvae did not survive in freshwater.

• Journal of Aquaculture
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• v.6 no.4
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• pp.235-253
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• 1993

The freshwater crab, Eriocheir japonicus inhabits from sub-tropical to temperate zone in Asia. This species belongs to a large size group among freshwater crabs. Common size of this crab is 5-6cm in carapace length and occasionally 7cm in carapace length. This species of crab used to inhabit in estuaries, rivers and inland waters in Korea. However, natural population recently has been rapidly decreased because of pollution and lost their habitats by suburban development. Therefore, development of proper methods of seedling production to increase natural stock became necessity. As parts of achieving this goal, duration from mating to spawning, egg incubation period, and egg development of this species were studied. The influence of temperatures and salinities on the egg incubation and hatching was also investigated. It took 2-8 hours from mating to egg spawning and the spawning lasted 3-9 hours from the first spawning. Egg numbers per female (6cm in carapace length) were 380,000­410,000. Optimum temperature for egg incubation was $17\~23^{\circ}C$ and optimum salinity, $14.0\~31.5\%o$. Incubation period of the eggs at $14^{\circ}C,\;17^{\circ}C,\;20^{\circ}C,\;26^{\circ}C,\;and\;28^{\circ}C$ was 42, 28, 21, 15, and 14 days. respectively. Relation between temperature (X) and incubation days (Y) was LogY = Log 2764.267 - 1.608 LogX. A female can spawn 4-6 times per year by manipulation of environmental conditions. Under the conditions of $18^{\circ}C\;and\;24.5\%o$, it took 6 days up to embryo formation, 18 days up to compound eye formation, 22 days up to abdominal movement, and 25 days up to hatch out as zoea larvae.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.3 no.1
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• pp.52-64
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• 1970

Fingerlings hatched from the eggs of the puffer, Fugu rubripes, which were spawned on May 21, 1969, and were cultivated. The results of their growth during 150 days, until October 25th, are summarized as follows: 1. The eggs began to hatch after 163 hours, at a water temperature of 15.9 to $17.4^{\circ}C$, and the hatching rate was $61.56\%$. 2. They reached the post-larval stage 6 to 7 days after hatching, and at this time a high mortality occurred. The mortality rate was 57.26 to $68.0\%$. 3. Sixteen days after hatching some of the larger fingerlings (6.7mm in total length) began attacking some of the smaller ones (4.6mm in to total length). 4. Twenty five to twenty eight days after hatching, the fish changed their food, and at this time a second high mortality occured. The total mortality rate amounted to 90.7 to $90.9\%$ of the total hatch. 5. After the fingerling stage. cannibalism occurs. The fish usually attack the caudal part of other fingerlings. It occurs regardless of body length and when the food supply is short. 6. The food coeffiicient at the age of 46 days (when body length is 53 to 68 mm) was 5.5 for short-necked clams, 8.5 for earth-worms, and 8.7 for fishes. 7. A third hish mortality occurred 53 to 63 days after hatching, the total mortality amounting to 95.76 to $97.34\%$, and the main cause of the mortality was found to be rickets resulting from nutritional deficiency 8. The growth rates were as follows: 2.68mm just after hatching, 3.84mm at the age of 10 days; 7.96mm after 25 days; 20.96mm or 130mg after 40 days; 73.68mm or 9.06g after 80 days: and 123mm or 31.8g after 150 days. 9. The water temperature during the above period was 15.7 to $28.4^{\circ}C$ with an average of $22.10^{\circ}C$ and the salinity was 25.53 to $34.50\%$ with an average of $32.07\%$, 10. The young of this species could endure well a wide range and sudden rise in salinity, and could survive easily when the salinity suddenly fell to $5\%$, but a considerable mortality occured when it fell to $3\%$. 11. When the fish were tranferred directly to fresh water from normal sea water they died out in 9 hours and 40minutes. However, when transferred from water of $5\%$ salinity at which they were reared for 54 days, they survived for 60 hours and 40 mimutes longer than in the former case.

• Journal of Embryo Transfer
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• v.29 no.3
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• pp.265-271
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• 2014

Implementation of smart embryo technologies in cattle e.g. ovum pick-up followed by in vitro embryo production (OPU-IVP). Seasonal variation is important factor for follicular growth, oocytes quality, quantity and developmental competence. Therefore the aim of present study was carried out to investigated whether the seasons (hot and cool) effect on follicular development, oocyte recovery and subsequent embryo development. Follicular oocytes were aspirated from Korean native cows (Hanwoo) by the ovum pick-up (OPU) method, which was performed 24 times during two different seasons, the hot (July to September) and cool (October to December), from OPU donors. The recovered oocytes were classified according to morphological categories and used for in vitro embryo production (IVEP). The mean number of total follicles was significantly higher (p<0.05) during the hot season ($18.32{\pm}2.26$) compared to cool season ($15.41{\pm}3.34$). Furthermore, seasons did not significantly effect on the number of oocytes recovered (hot season: 41.16% vs. cool season: 46.14%). However, the average number of Grade A oocytes was significantly greater during hot ($1.75{\pm}1.86$) season compared to the cool season ($1.00{\pm}1.46$), but there was no significant difference of other grades oocytes. The cleavage rate (hot: 66.67% vs. cool: 63.3%) and embryo development (hot: 58.95% vs. cool: 56.97%) did not differ significantly between the seasons. In conclusion, the results of present study suggest that the season (hot and cool) does not have effects on the oocyte recovery and embryo developmental competence of in vitro cultured embryos.

• Korean Journal of Poultry Science
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• v.17 no.3
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• pp.167-177
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• 1990

In order to investigate an effect of the feed restriction on the laying performance, 208 White Leghorn strain layer of 36 week-old were divided to 4 groups of 52 birds each and raised for 1 week of previous feeding and for the subsequent 3 weeks of experimental restricted feeding. The egg production, daily egg mass and feed efficiency of four groups of birds fed daily 100g, 95g, 90f and 85g of a commercial diet, respectively, which were compared with those of the original 2879 birds fed l13g of diet per day as control. During 3 weeks of experimental restricted feeding, period, daily body weight nam was decreased linearly as the degree of restriction increased in birds fed 100, 95, 90 and 85g of diet. Hen day egg production, egg weight and daily egg mass was linearly related to the diet intake level. then feed intake(x, g day$^{-1}$ ) showed a positive regression equations with the henday egg production(y, % ), egg weight(y, g egg$^{-1}$ ) and egg mass(y, g bird$^{-1}$ ) as y＝38.75＋0.3753$\times$(r＝0.503, n＝15), y＝48.2＋0.08868$\times$(r＝0.835, n＝ 15) and y＝15.69＋0.2786$\times$(r＝0.597, n＝15), respectively. Feed efficiency was increased to a plateau in birds fed 95g of diet. The estimated energy utilization for egg production was reached to a plateau in birds fed 95g of diet and the highst protein utilization was shown in birds fed 90g of diet anions birds fed graded levels of diet. And the feed restriction did not affect on the egg shell contents, while protein contents of egg were shown a trend to be increased and lipids and cholesterol contents of eggs was decreased according to the diet intake lowered. The results suggested that the improved feed efficiencies of birds restricted under 16% of diet(above 95g of diet) will be due to increased energy and protein utilization for egg production and feed restriction above 16% will be aboided. In the range from 113g to 95g of diet feeding, the crude profit was increased as the feed restricted in the case of egg price 600 won kg$^{-1}$ and feed price 200 won kg$^{-1}$ .

• Journal of Sericultural and Entomological Science
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• v.50 no.2
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• pp.63-70
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• 2012

This study was conducted to investigated the distribution, ecological character and life cycle of Black Soldier Fly(BSF), Hermetia illucens, to determine effects of BSF on composing ability to the livestock feces. The distribution of BSF was defined in all parts of the country in Korea. Its main habitat was found to be areas near cattle sheds, feces sheds, living waste dump grounds, and food waste dump grounds. Excellent species of the bio-conversion using the livestock feces were selected out of two kinds of solider flies(Hermetia illucens and Ptecticus tenebrifer). And effects of BSF on composing ability to the livestock feces were pig feces 0.5 g, cattle feces 0.43 g and chicken feces 0.3 g per a larvae of BSF. In the examination of the ability of BSF to decompose food waste, volume of the livestock feces decreased by weight pig feces 77, cattle feces 83.62 and chicken feces 80%. Observed characteristics of BSF larvae and pupae stage were by developmental stage may be summarized as follows: size and length of larvae and pupae of BSF were higher than in pig feces and mixed sawdust treatments. In order to develop artificial indoor rearing techniques, The pupation and emergence rate were not visible the difference but important factor of the mating and egg-laying was a sun light. And it was visible the difference in mating ratio. Mating rate of H. illucens by seasonal change was the highest from May to July. Mating mostly occurred between 10 : 00 and 12 : 00 during which light intensity is high. As the egg-laying medium for artificial egg collection, calf feed and food waste were most effective. The ratio of egg-laying medium is waste food : sawdust(1 : 1) in volume. For egg-laying materials, flower foam and wood with holes were used; holes sized 3 ~ 5 mm in diameter, 7 ~ 10 mm in depth were most preferred for egg-laying.

• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.167-176
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• 1997

This study was carried out to evaluate the potential for preselection of transgenic embryos prior to transfer into recipient animals. In these experiments, I used a 3.2 kb transgene which contained the neomycin resistance gene (neo) and lac Z gene driven by the $\beta$ actin promoter (iacZ Ineo). Oocytes were aspirated from abattoir ovaries, matured in TCM-199 supplemented with 10% fetal bovine serum (FBS), 5 ${\mu}\textrm{g}$/ml LH, 0.5 ${\mu}\textrm{g}$/ml FSH, 100 unit/ml penicillin, and 100 ${\mu}\textrm{g}$/ml streptomycin for 22 to 24 hrs then inseminated with a sperm suspension of 1 X 10$^6$ sperm/ml containing 5 ${\mu}\textrm{g}$/ml of heparin. At 18-20 hrs after insemination, cumulus cells were removed by vortexing and pronuclei of centrifuged zygotes microinjected with the lacZ/neo construct (3 ng/$\mu$l). All cultures were carried out in CR1aa with transfected BRL monolayers containing 3 mg/ml BSA, 20 $\mu$/ml NEM amino acids and 40 $\mu$I/ml BME amino acids. To identify the appropriate concentration of G418 for selection, non-microinjected zygotes were cultured in the presence of 0, 50, 100 and 200 $\mu$g/ml of G418. After 8 days of culture in these treatments, 44/145 (30.3%), 13/150 (8. 7%), 1/151 (0.7%) and 0/134 (0.0%) devel-oped to the blastocyst stage in 0, 50, 100 and 200 $\mu$g/ml of G418, respectively. A total of 1,127 zygotes were microinjected and placed into culture (without G418) and subsequently 710 (63.0%) cleaved. At 48 hrs post-injection, embryos ($\geq$2-cell) were randomly assigned to control (medium alone) or G418 (100 ${\mu}\textrm{g}$/ml) treatments. A control culture of 740 non-microinjected embryos from the same replicates of embryos were also placed into control medium. After 8 days in culture, 54/343 (15.7%) and 22/367 (6.0 %) of the microinjected embryos developed to the blastocyst stage in control and G418 media, respectively. A total of 151/740 (27.2%) of the non-microinjected embryos placed in the control medium developed to the blastocyst stage. The blastocysts in the control treatment had a mean of 70.7 ${\pm}$ 4.7 cells of which 23.1 ${\pm}$ 2.6 (32.7%) stained for $\beta$-Gal activity. B1astocysts in the G418 treatment had a mean of 48.8${\pm}$7.5 cells of which 40.3 ${\pm}$ 4.1 (82.6%) stained for $\beta$-Gal ac tivity (P<0.05). In the control treatment 26 of 30 (87.0%) blastocysts had some cells with $\beta$-Gal activity while all of the blastocysts in the G418 treatment had some cell with $\beta$-Gal ac tivity (P<0.05). However, ICM colonies in either control or G418 treatments were not expressed any epiblast cell except of trophetoderm celIs. The $\beta$-actin promoter/lacZ gene may not be e expression or silence expression in epiblast cells These results clearly show an enrichment of blastocysts expressing the transgene in the majority of their cells after culture in the presence of G418. The exogeneous gene was not express a and silence in ICM colonies especiallly epiblast cells except of trophectederm cells. Even though the higher rate cell number expressed of exogeneous gene in the G418 treatments, a total cell number was decrease significantly (P<0.05) of which might be also drop of the establishment of ES like-cell colonies and production of transgenic animals. However, futher studies need to determine the viability of these selected embryos and the avability of production of transgenic offspring.

• Journal of Sericultural and Entomological Science
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• v.52 no.2
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• pp.110-116
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• 2014

Recently the number of the butterflies, Mellicta ambigua, had been decreasing rapidly, and already disappeared at many habitat. In this studies, we investigated ecological environment of Mellicta ambigua for preparing of primary research data recovering habitat, and studied on bionomical characteristics. Two different habitat, Jindo and Inje, were selected for investigation of ecological environment. We investigated four times during 3-month, from June to August in 2012. In Jindo, we observed more than 100 butterflies and a lot of host plants, Melampyrum roseum var. japonicum. But only 5 butterflies and only a few host plants, Veronicastrum sibiricum were observed in Inje. We could not observe the eggs, the larva and pupa on the host plants at all. For finding of bionomical characteritics, we reared butterflies at natural conditions. Collected 3-female butterflies from Jindo laid 465 eggs on the leaves of 3-host plants, Veronicastrum sibiricum. 120 ~ 186 eggs per each female were laid in the shape of cluster. An egg was globular shape, 0.6 mm diameter and 0.7 mm height. The egg periods were $9.96{\pm}0.4days$ after ovipositioning, and the hatchability was 95.% at natural condition. The larval periods were $4.1{\pm}0.6days$ (1st instar), $2.1{\pm}1.0days$ (2nd), $8.1{\pm}0.7days$ (3rd), $239.2{\pm}10.9days$ (4th), $12.3{\pm}1.3days$ (5th), $17.1{\pm}1.1days$ (6th), $10.5{\pm}1.0days$ (7th) each other. The larva of 4th instar overwintered in the nest that had been made into the leaf of host plant with secreted thread as a group until early March next year. In the early March next year, overwintered larva went around their nest in search of host plants, and went to other host plants, Veronica persica and Plantago asiatica, sometimes. The overwintered larva of Mellicta ambigua could grow up on two other host plants normally. In the following experiment, the butterflies of Mellicta ambigua laid eggs on the leaves of Plantago asiatica, but the 1st instar larva from eggs died all. The headwidth of each developmental larval stage were $0.28{\pm}0.02mm$ (1st), $0.45{\pm}0.02mm$ (2nd), $0.58{\pm}0.02mm$ (3rd), $0.75{\pm}0.03mm$ (4th), $0.89{\pm}0.05mm$ (5th), $1.23{\pm}0.06mm$ (6th), $2.13{\pm}0.11mm$ (7th). The pupal ratio was 92.0%. The pupal period were $9.1{\pm}1.6days$, and the emergence rate was 88.6%. As a result we determined that Mellicta ambigua can rear at natural conditions. But indoor-rearing is considered to be difficult and not useful industrially, because they have long term larval stage and only one life cycle per an year.