Prediction of canine ovulation using serum estradiol concentration

Zhao Minghui,Lee Seunghoon,No Jingu,Nam Yoonseok,Jeong Hae-yun,Ock Sun A,Yun JeongHee,Kim Dong-Hoon,Tai-Young 한국수정란이식학회 2017 한국수정란이식학회 학술대회 Vol.2017 No.05

Canine cloning have been succeeded for a decade. To obtain in vivo matured dog oocytes, Serum progesterone (P4) level were employed for ovulate determination. However, accuracy of P4 methods is not satisfied. The aim of this study was to compare both methods of serum estradiol (E2) and P4 on the accuracy of canine ovulation determination. Canine serum P4 and E2 concentration during both proestrus and estrus were detected. Correlation between accuracy of each method and environment temperature were analyzed. Following ovulation, oocytes were collected by surgery. As a result, higher percentage of mature oocytes was obtained when using E2 (56.43%) as compared to P4 (39.60%). Accuracy of P4 increased from spring (30.76%) to summer (47.92%) and decreased in autumn (37.50%) and winter (29.16%) gradually. Especially, E2 maintained about 50% to 65% whatever the season and temperature. Correlation analyze showed that dynamic of P4 accuracy highly correlated with environment temperate (Rp4=0.862) but E2 could not be affected by the temperature (RE2=0.199). To determine whether obtained oocytes by E2 method could be used for canine cloning, twenty canines were selected as oocyte donors, and two puppies were produced after somatic cell nuclear transfer(SCNT) and embryo transfer(ET) with the oocytes by E2 method. In conclusion, comparing to the P4 method, the E2 is an accuracy and reliable method for canine cloning.

Dog cloning with in vivo matured oocytes obtained using electric chemilumiescence immunoassay-predicted ovulation method

Lee Seunghoon,Zhao Minghui,No Jingu,Nam Yoonseok,Jeong Hae-yun,Ock Sun A,Yun JeongHee,Kim Dong-Hoon,Hur Tai-Young 한국수정란이식학회 2017 한국수정란이식학회 학술대회 Vol.2017 No.05

To obtain in vivo matured oocytes for dog cloning, serum progesterone (P4) level were employed for ovulate determination. Radioactive immunoassay (RIA) is a traditional serum hormone assay method with highly radioactivity. The aim of this study was to evaluate the reliability of RIA and to compare its canine serum P4 concentration determination accuracy to that of the electric chemiluminescence immunoassay (ECLI). To obtain in vivo matured oocytes for canine somatic cell nuclear transfer, serum P4 levels were accurately measured with both methods of RIA and ECLI. Although both methods detected similar P4 level before ovulation, the mean P4 concentration using ECLI was significantly higher than that using RIA from 3days before ovulation. Following ovulation, oocytes were collected by surgery, and a lower percentage of mature oocytes were observed using ECLI (39%) as compared to RIA (67%) if 4-8ng/ml of P4 were criteria for determination of ovulation. On other hand, high percentage of mature oocytes was observed using ECLI when 6–15 ng/mL of progesterone was criteria for ovulation determination. To determine whether in vivo oocytes obtained by ECLI method could be used for canine cloning, six canines were selected as oocyte donors and two puppies were produced after SCNT and embryo transfer. In conclusion, compared to the traditional RIA method, the ECLI method is a safe and reliable method for canine cloning.

Comparing Gut Microbial Composition and Functional Adaptations between SPF and Non-SPF Pigs

( Haesun Lee ),( Woncheoul Park ),( Jingu No ),( Nam Woong Hyung ),( Ju-yeong Lee ),( Seokho Kim ),( Hyeon Yang ),( Poongyeon Lee ),( Eunju Kim ),( Keon Bong Oh ),( Jae Gyu Yoo ),( Seunghoon Lee ) 한국미생물생명공학회 2024 Journal of microbiology and biotechnology Vol.34 No.7

The gut microbiota is a key factor significantly impacting host health by influencing metabolism and immune function. Its composition can be altered by genetic factors, as well as environmental factors such as the host's surroundings, diet, and antibiotic usage. This study aims to examine how the characteristics of the gut microbiota in pigs, used as source animals for xenotransplantation, vary depending on their rearing environment. We compared the diversity and composition of gut microbiota in fecal samples from pigs raised in specific pathogen-free (SPF) and conventional (non- SPF) facilities. The 16S RNA metagenome sequencing results revealed that pigs raised in non-SPF facilities exhibited greater gut microbiota diversity compared to those in SPF facilities. Genera such as Streptococcus and Ruminococcus were more abundant in SPF pigs compared to non-SPF pigs, while Blautia, Bacteroides, and Roseburia were only observed in SPF pigs. Conversely, Prevotella was exclusively present in non-SPF pigs. It was predicted that SPF pigs would show higher levels of processes related to carbohydrate and nucleotide metabolism, and environmental information processing. On the other hand, energy and lipid metabolism, as well as processes associated with genetic information, cell communication, and diseases, were predicted to be more active in the gut microbiota of non-SPF pigs. This study provides insights into how the presence or absence of microorganisms, including pathogens, in pig-rearing facilities affects the composition and function of the pigs' gut microbiota. Furthermore, this serves as a reference for tracing whether xenotransplantation source pigs were maintained in a pathogen-controlled environment.

Dog cloning with <i>in vivo</i> matured oocytes obtaining using serum estradiol levels for predicting time of ovulation

Zhao, Minghui,Lee, Seunghoon,Kim, Dong-Hoon,No, Jingu,Nam, Yoonseok,Ock, Sun A.,Ko, Yeoung-Gyu,Hur, Tai-Young Elsevier 2018 Theriogenology Vol.107 No.-

<P><B>Abstract</B></P> <P>Dog cloning using <I>in vivo</I>-matured oocytes has been carried out for a decade. To obtain mature oocytes, serum progesterone (P<SUB>4</SUB>) levels are used to evaluate ovulation. However, the accuracy of these methods is not sufficient. Thus, the aim of the present study was to verify the feasibility of serum estradiol (E<SUB>2</SUB>) on canine ovulation determination as assessed by the percentage of dogs yielding mature oocytes. <I>In vivo</I>-matured oocytes were utilized for canine somatic cell nuclear transfer (SCNT), and serum P<SUB>4</SUB> and E<SUB>2</SUB> levels were assessed to determine ovulation and oocyte maturation. Canine serum P<SUB>4</SUB> and E<SUB>2</SUB> concentrations during both pro-estrus and estrus were analyzed by electrochemiluminescence immunoassay. The percentage of dogs yielding mature oocytes using each of the two ovulation prediction methods were compared, and correlations between the percentage of each method and temperature were analyzed. Following evaluation, oocytes were collected surgically, and a significantly higher percentage (<I>P</I> < 0.05) of dogs yielding mature oocytes was observed using E<SUB>2</SUB> (56.43%) for ovulation detection as compared with that using P<SUB>4</SUB> (39.60%). The percentage of dogs yielding mature oocytes using P<SUB>4</SUB> significantly lower (<I>P</I> < 0.05) than E<SUB>2</SUB> in autumn (P<SUB>4</SUB>, 37.50% vs. E<SUB>2</SUB>, 52.00%) and winter (P<SUB>4</SUB>, 29.17% vs. E<SUB>2</SUB>, 59.09%). Using E<SUB>2</SUB>, the percentage was maintained at about 52.00–66.67% regardless of the season and temperature. Correlation analysis showed that the dynamic of percentage of dogs yielding mature oocyte using P<SUB>4</SUB> was highly correlated with environmental temperature (R<SUB>P4</SUB> = 0.862), whereas E<SUB>2</SUB> was not affected by temperature (R<SUB>E2</SUB> = 0.199). To determine whether serum E<SUB>2</SUB> could be used for ovulation prediction for canine cloning, ovulation of 25 and 19 dogs (<I>P</I> < 0.05) were predicted using P<SUB>4</SUB> or E<SUB>2</SUB> methods, respectively and two puppies, one from each ovulation prediction method, were obtained after SCNT and embryo transfer. Thus, compared with the P<SUB>4</SUB> method, E<SUB>2</SUB> was an accurate and reliable method for canine cloning.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The percentage of dogs yielding mature oocytes following prediction of ovulation using serum P<SUB>4</SUB> was affected by temperature. </LI> <LI> The percentage of dogs yielding mature oocytes following estimation of ovulation with serum E<SUB>2</SUB> was higher than that of dogs with oocytes recovered after the use of P<SUB>4</SUB> for ovulation determination. </LI> <LI> Cloned pups were produced when serum E<SUB>2</SUB> was used for predicting ovulation time. </LI> </UL> </P>

Isolation and characterization of cultured chicken oviduct epithelial cells and in vitro validation of constructed ovalbumin promoter in these cells

Yang, Hyeon,Lee, Bo Ram,Lee, Hwi-Cheul,Jung, Sun Keun,Kim, Ji-Youn,No, Jingu,Shanmugam, Sureshkumar,Jo, Yong Jin,Lee, Haesun,Hwang, Seongsoo,Byun, Sung June Asian Australasian Association of Animal Productio 2021 Animal Bioscience Vol.34 No.8

Objective: Transgenic hens hold a great promise to produce various valuable proteins. Through virus transduction into stage X embryo, the transgene expression under the control of constructed chicken ovalbumin promoters has been successfully achieved. However, a validation system that can evaluate differently developed ovalbumin promoters in in vitro, remains to be developed. Methods: In the present study, chicken oviduct epithelial cells (cOECs) were isolated from oviduct tissue and shortly cultured with keratinocyte complete medium supplemented with chicken serum. The isolated cells were characterized with immunofluorescence, western blot, and flow cytometry using oviduct-specific marker. Chicken mutated ovalbumin promoter (Mut-4.4-kb-pOV) was validated in these cells using luciferase reporter analysis. Results: The isolated cOECs revealed that the oviduct-specific marker, ovalbumin protein, was clearly detected by immunofluorescence, western blot, and flow cytometry analysis revealed that approximately 79.40% of the cells contained this protein. Also, luciferase reporter analysis showed that the constructed Mut-4.4-kb-pOV exhibited 7.1-fold (p<0.001) higher activity in the cOECs. Conclusion: Collectively, these results demonstrate the efficient isolation and characterization of cOECs and validate the activity of the constructed ovalbumin promoter in the cultured cOECs. The in vitro validation of the recombinant promoter activity in cOECs can facilitate the production of efficient transgenic chickens for potential use as bioreactors.