• Journal of Animal Reproduction and Biotechnology
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• v.37 no.2
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• pp.80-86
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• 2022

The ability to determine the sex of bovine embryos before the transfer is advantageous in livestock management, especially in dairy production, where female calves are preferred in milk industry. The milk production of female and male cattle benefits both the dairy and beef industries. Pre-implantation sexing of embryos also helps with embryo transfer success. There are two approaches for sexing bovine embryos in farm animals: invasive and non-invasive. A non-invasive method of embryo sexing retains the embryo's autonomy and, as a result, is less likely to impair the embryo's ability to move and implant successfully. There are lists of non-invasive embryo sexing such as; Detection of H-Y antigens, X-linked enzymes, and sexing based on embryo cleavage and development. Since it protects the embryo's autonomy, the non-invasive procedure is considered to be the safest. Invasive methods affect an embryo's integrity and are likely to damage the embryo's chances of successful transformation. There are different types of invasive methods such as polymerase chain reaction, detection of male chromatin Y chromosome-specific DNA probes, Loop-mediated isothermal amplification (LAMP), cytological karyotyping, and immunofluorescence (FISH). The PCR approach is highly sensitive, precise, and effective as compared to invasive methods of farm animal embryonic sexing. Invasive procedures, such as cytological karyotyping, have high accuracy but are impractical in the field due to embryonic effectiveness concerns. This technology can be applicable especially in the dairy and beef industry by producing female and male animals respectively. Enhancing selection accuracy and decreasing the multiple ovulation embryo transfer costs.

• Korean Journal of Poultry Science
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• v.48 no.1
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• pp.41-50
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• 2021

Although feather-sexing using sex-linked genes related to feather development is a widely used chick sexing method in the poultry industry, the feather-sexing method has yet to be used for Korean native chickens (KNCs). The purpose of this study was to construct a KNC feather-sexing line using early-feathering (EF) and late-feathering (LF) genes for industrial application. Using 557 reddish-brown KNCs as the basal flock, frequencies of the EF (k) and LF (K) genes were estimated to be 0.814 and 0.186, respectively. This indicating that it would be feasible to construct a feather-sexing line using this chicken group, and we accordingly constructed EF paternal and LF maternal lines. On the basis of test-cross for the selection of LF homozygous (KK) males in the maternal line, we confirmed that three of 40 chickens were homozygous males. The survival rate, body weight, days at first egg-laying, hen-day egg production, and egg weight were analyzed to compare the production performance of EF and LF chickens. The results revealed that EF chickens were characterized by a superior survival rate, whereas LF chickens were superior in terms of egg production rate. However, no differences between LF and EF chickens were detected with respect to other production performance parameters. In addition, assessment of the fitness of sexed chicks produced in the established KNC feather-sexing lines revealed that the accuracy of sexing was 98.6%. Collectively, these findings indicate the feasibility of constructing effective KNC feather-sexing lines with potential industrial application.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2003.10a
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• pp.92-92
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• 2003

• Korean Journal of Poultry Science
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• v.40 no.3
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• pp.263-270
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• 2013

The vent sexing and the auto-sexing by using sex-linked traits are general sexing methods of day-old chicks. Currently, the feather sexing which is based on the differences in the feather characteristics at hatching is the representative sexing method of chicken, because the late-feathering is sex-linked trait. The feather sexing can be used if the breed has dominant feathering gene (K) in maternal and recessive gene ($k^+$) in paternal. Therefore it is necessary to identify the association of feathering genes and quantitative traits in chickens. In this study, we investigated the influence of the rate of feathering on productive traits in Korean Native Chicken. In results, there was no significant difference between early-feathering chickens and late-feathering chickens in reproductive performance such as fertility and hatchability. Livability, body weights, egg production, egg weight and egg quality also did not significantly differ between early- and late-feathering chickens. Age at first egg was the only trait of those tested in which significant difference was observed. The early-feathering chickens laid eggs 3 days earlier than late-feathering chicken. As a result, there is no influence of feathering phenotypes on productive performance in Korean Native Chickens. Consequentially, establishing the feather sexing strain is available using the Korean Native Chicken breed without considering of the effect of feathering genes on productive traits.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.199-205
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• 2013

The present study was conducted to compare on embryo survival rates by blastomere isolation methods, and establish the optimal PCR procedure for perform the sexing of bovine blastocysts produced by IVF. IVF embryos used in the study was used the Bisected or Sliced methods for blastomere isolation, and the survival rates of blastocyst with rapid way of sexing PCR was assessed. In the present study for survival rates in blastocyst was the total cleavage rate was 75% and a blastocyst development among cleaved embryos was 40%. Survival rate of embryos treated with intact, bisected or sliced method was 100, 63.3 or 81.3%, respectively. Therefore, survival rate of embryos treated with sliced method was higher compared to that of embryos treated with bisected method. The sexing rate of female or male was not significantly different between S4BFBR primer and BSY + BSP primer (1.75 : 1 vs. 1.43 : 1), respectively. Because of the PCR amplification using the S4BFBR primer was simpler method than multiplex PCR amplification method. Furthermore, the accuracy of sexing rate and reduction of PCR work time between 2-step and 3-step of PCR methods was 98.0% / 1.5 hr and 97.0% / 3.5 hr, respectively. Based on these results, it can be suggested that the sliced and PCR methods we developed was very effective method to reduce time consuming and procedure of PCR amplification for sexing with the increase of survival rate on the blastocyst.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.92-92
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• 2003

Accurate analysis of nuclear status is needed when biopsied-blastomeres are used for embryo sexing. In this study, the nuclear status of blastomeres derived from 8- to 16-cell stage IVF bovine embryos was analyzed to evaluate the representative of single blastomere for embryo sexing. When 55 embryos were analyzed by PCR following biopsy, the coincident rate of sex determination between biopsied-single blastomere and matched blastocyst by PCR was 80 %. Karyotyping of biastomeres in 8- 16-cell stage bovine embryos was conducted to assess chromosome status of IVF embryos. To establish karyotyping of blastomeres, concentrations of vinblastine sulfate and duration of exposure time for metaphase plate induction with 8- to 16-cell stage bovine embryos were tested. The most effective condition for induction of metaphase plate (>45%) was 1.0 ug/ml vinblastine sulfate treatment for 15 h. In 22 embryos under the condition, only 8 embryos out of ten that had a normal diploid chromosome complement showed a sex-chromosomal composition of XX or XY (36.4%) and 2 diploid embryos showed mosaicism of the opposite sex of XX and XY in blastomeres of embryo (9.1%). One haploid embryo contained only one X-chromosome (4.5%). Four out of the other 11 embryos having a mixoploid chromosomal complement contained haploid blastomere with wrong sex chromosome (18.2%). These results suggested that morphologically normal bovine embryos derived from IVF had considerable proportion of mixoploid and sex-chromosomal mosaicism which could be the cause of discrepancies of the sex between biopsied-single blastomere and matched blastocyst by PCR analysis.

• Korean Journal of Poultry Science
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• v.46 no.4
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• pp.279-286
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• 2019

Currently, feather-sexing, which is based on differences in feather development at hatching, is a widely used chick sexing method in the poultry industry. For effective chicken feather-sexing, paternal early-feathering (EF) chickens and maternal late-feathering (LF) chickens must be bred. Therefore, it is critical to identify the effect of EF and LF patterns on production traits in chickens. Thus, the purpose of this study is to analyze and compare the production performances between 522 EF and 232 LF chickens in order to establish the Korean native chicken feather-sexing lines. The results showed that the survival rate of the LF group was significantly higher than that of the EF group from hatching to 52 weeks of age (P<0.05). Body weight, however, was not significantly different between the two groups at all ages. LF and EF groups did not significantly differ in age at first egg laying. However, the hen-day and hen-housed egg production of the LF group were significantly higher than those of EF group (P<0.01). No significant differences were found between the EF and LF groups in all egg quality indicators such as egg weight, eggshell color, albumin height and Haugh unit. Because the breeding target of Korean native commercial chicken is meat-type chicken, feather-sexing strains of Korean native chicken should be established using weighing-based paternal EF lines and laying-based maternal LF lines. Therefore, these results are critical for establishing desirable and effective feather-sexing strains.

• Journal of Animal Science and Technology
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• v.54 no.4
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• pp.267-274
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• 2012

The method of sexing based on differences in the rate of feather growth provides a convenient and inexpensive approach. The locus of feather development gene (K) is located on the Z chromosome and can be utilized to produce phenotypes that distinguish between the sexes of chicks at hatching. To establish the auto-sexing native chicken strains, this study analyzed the genotype frequency of the feathering in domestic chicken breeds. The method of classification of slow- and rapid-feathering chickens was also investigated. In the slow-feathering chicks, the coverts were either the same length or longer than the primary wing feathers at hatching. However, the rapid-feathering chicks had the primary wing feathers that were longer than the coverts. The growth pattern of tail feather also distinctively differed between the rapid- and slow-feathering chicks after 5-days. The accuracy of wing feather sexing was about 98% compared with tail sexing. In domestic chicken breeds, Korean Black Cornish, Korean Rhode Island Red, and Korean Native Chicken-Red had both dominant (K) and recessive ($k^+$) feathering genes. The other breeds of chickens, Korean Brown Cornish, Ogol, White Leghorn, Korean Native Chicken-Yellow, -Gray, -White and -Black had only the recessive feathering gene ($k^+$). Consequently, feather sexing is available using the domestic chicken breeds. Establishing the maternal stock with dominant gene (K-) and paternal stock with recessive gene ($k^+k^+$), the slow-feathering characteristic is passed from mothers to their sons, and the rapid-feathering characteristic is inherited by daughters from their fathers.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.11
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• pp.1531-1535
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• 2002

It has been known that the sex of chicken cells can be most accurately identified by fluorescence in situ hybridization (FISH). However, the presently available FISH has not been widely used for sex identification, because the procedures for cell preparation and FISH itself are complicated and time-consuming. The present study was undertaken to test a rapid FISH procedure for sexing chicken. A FISH probe was simultaneously synthesized and labeled with digoxigenin by polymerase chain reaction (PCR) targeting a 416 bp segment of the 717 bp XhoI family fragment which is repeated over 10 thousand times exclusively in the W chromosome. Sexing by FISH was performed on cytological preparations of early embryos, adult lymphocytes and feather pulps of newly hatched chicks. The DNA probe hybridized to all types of uncultured interphase as well as metaphase female but not male cells that had been examined. Moreover, consistent with the known site of the XhoI family, the hybridization signal was localized to the pericentromeric region of the W chromosome. We, therefore, conclude that the present PCR-based FISH can be used as a rapid and reliable sex identification procedure for chicken.

• 대한생식의학회:학술대회논문집
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• 1995.12a
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• pp.43-45
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• 1995