Background Hexafluoropropylene oxide dimer acid(GenX),a substitute for per-and polyfluoroalkyl substances,has been widely detected in various environmental matrices and foods recently,attracting great attention.Howeve...Background Hexafluoropropylene oxide dimer acid(GenX),a substitute for per-and polyfluoroalkyl substances,has been widely detected in various environmental matrices and foods recently,attracting great attention.However,a systematic characterization of its reproductive toxicity is still missing.This study aims to explore the male reproductive toxicity caused by GenX exposure and the potential cellular and molecular regulatory mechanisms behind it.Results Normally developing mice were exposed to GenX,and testicular tissue was subsequently analyzed and validated using single-cell RNA sequencing.Our results revealed that GenX induced severe testicular damage,disrupted the balance between undifferentiated and differentiated spermatogonial stem cells,and led to strong variation in the cellular dynamics of spermatogenesis.Furthermore,GenX exposure caused global upregulation of testicular somatic cellular inflammatory responses,increased abnormal macrophage differentiation,and attenuated fibroblast adhesion,disorganizing the somatic-germline interactions.Conclusions In conclusion,this study revealed complex cellular dynamics and transcriptome changes in mouse testis after GenX exposure,providing a valuable resource for understanding its reproductive toxicity.展开更多
Background Pork quality can directly affect customer purchase tendency and meat quality traits have become valu-able in modern pork production.However,genetic improvement has been slow due to high phenotyping costs.In...Background Pork quality can directly affect customer purchase tendency and meat quality traits have become valu-able in modern pork production.However,genetic improvement has been slow due to high phenotyping costs.In this study,whole genome sequence(WGS)data was used to evaluate the prediction accuracy of genomic best linear unbiased prediction(GBLUP)for meat quality in large-scale crossbred commercial pigs.Results We produced WGS data(18,695,907 SNPs and 2,106,902 INDELs exceed quality control)from 1,469 sequenced Duroc×(Landrace×Yorkshire)pigs and developed a reference panel for meat quality including meat color score,marbling score,L*(lightness),a*(redness),and b*(yellowness)of genomic prediction.The prediction accuracy was defined as the Pearson correlation coefficient between adjusted phenotypes and genomic estimated breeding values in the validation population.Using different marker density panels derived from WGS data,accuracy differed substantially among meat quality traits,varied from 0.08 to 0.47.Results showed that MultiBLUP outperform GBLUP and yielded accuracy increases ranging from 17.39%to 75%.We optimized the marker density and found medium-and high-density marker panels are beneficial for the estimation of heritability for meat quality.Moreover,we conducted genotype imputation from 50K chip to WGS level in the same population and found average concord-ance rate to exceed 95%and r^(2)=0.81.Conclusions Overall,estimation of heritability for meat quality traits can benefit from the use of WGS data.This study showed the superiority of using WGS data to genetically improve pork quality in genomic prediction.展开更多
Background The establishment of a robust gut microbiota in piglets during their early developmental stage holds the potential for long-term advantageous effects.However,the optimal timeframe for introducing probiotics...Background The establishment of a robust gut microbiota in piglets during their early developmental stage holds the potential for long-term advantageous effects.However,the optimal timeframe for introducing probiotics to achieve this outcome remains uncertain.Results In the context of this investigation,we conducted a longitudinal assessment of the fecal microbiota of 63 piglets at three distinct pre-weaning time points.Simultaneously,we gathered vaginal and fecal samples from 23 sows.Employing 16S rRNA gene and metagenomic sequencing methodologies,we conducted a comprehensive analysis of the fluctuation patterns in microbial composition,functional capacity,interaction networks,and colonization resistance within the gut microbiota of piglets.As the piglets progressed in age,discernible modifications in intestinal microbial diversity,composition,and function were observed.A source-tracking analysis unveiled the pivotal role of fecal and vaginal microbiota derived from sows in populating the gut microbiota of neonatal piglets.By D21,the microbial interaction network displayed a more concise and efficient configuration,accompanied by enhanced colonization resistance relative to the other two time points.Moreover,we identified three strains of Ruminococcus sp.at D10 as potential candidates for improving piglets’weight gain during the weaning phase.Conclusions The findings of this study propose that D10 represents the most opportune juncture for the introduction of external probiotic interventions during the early stages of piglet development.This investigation augments our comprehension of the microbiota dynamics in early-life of piglets and offers valuable insights for guiding forthcoming probiotic interventions.展开更多
Aflatoxin B_1(AFB_1) is one of the most potent mycotoxins affecting human health and animal production.To deeply understand the hosttoxin interaction,we performed CRISPR screening and identified cystathionine β-synth...Aflatoxin B_1(AFB_1) is one of the most potent mycotoxins affecting human health and animal production.To deeply understand the hosttoxin interaction,we performed CRISPR screening and identified cystathionine β-synthase(CBS) as a critical host gene affecting AFB_1 cytotoxicity.Mechanistic studies revealed that CBS affects AFB_1-induced cell death by regulating the abundance of protein post-translational modifications(PTMs) in host cells.First,AFB_1 disrupted the transfer of S-adenosylmethionine(SAM) from the cytoplasm to the mitochondria,thereby reducing the intra-mitochondrial protein methylation level.Deficient intra-mitochondrial protein methylation impaired mitochondrial function and caused cell death.CBS knockout(KO) can enhance SAM generation and mobilization to restore intramitochondrial SAM levels by rescuing the perturbed methionine cycle after AFB_1 exposure,thereby alleviating AFB_1-induced cell death.Second,AFB_1 decreased global protein ubiquitination levels by affecting gene expression of ubiquitin-modified enzymes.CBS-KO and pharmaceutical treatment correcting gene expression of ubiquitin-modified enzymes can rescue AFB_1-induced cell death.We also investigated two PTM-regulating small molecules,SAM and PR-619,which can increase cell viability in AFB_1-exposed cells.展开更多
Pig body measurement is an important evaluation criterion for breeding and production management.Automatic measurement algorithms for pig body sizes exhibit sensitivity to the point cloud posture,but non-standard pig ...Pig body measurement is an important evaluation criterion for breeding and production management.Automatic measurement algorithms for pig body sizes exhibit sensitivity to the point cloud posture,but non-standard pig postures may result in inaccurate joint point localization in body measurement,further affecting measurement accuracy and the commercial application of these algorithms.To address this challenge,this paper proposed a pig point cloud posture transformation method based on pig’s skeleton model to adjust non-standard postures before conducting body size measurements.The method utilized an improved L1-median skeleton model to extract the three-dimensional skeleton of the pig point cloud,capturing the skeleton joint points on the target pig’s head,body,and limbs.By binding the skeleton joint points with the local point cloud and using rotation matrices,non-standard postures were adjusted to standard ones,enabling accurate body size measurements.The experimental results demonstrated that the average relative errors between the transferred posture and the original standard posture were reduced to 0.89%in body length,0.76%in body width(front),1%in body width(back),0.89%in body height(front),1.7%in body height(back),2.03%in thoracic circumference,3.37%in abdominal circumference,and 1.89%in rump circumference.To conclude,the posture standardization transfer method can significantly reduce errors in important body size parameters such as body length,body height,and body width.The method displays a greater stability and robustness compared to existing posture normalization and regression adjustment methods,providing both guidance and insight for future research in intelligent agriculture.展开更多
Intramuscular fat(IMF) is a major meat-quality trait in pigs. The content of IMF is directly associated with the taste and flavor of pork. As a complex trait, there could be multiple genes affecting IMF content in por...Intramuscular fat(IMF) is a major meat-quality trait in pigs. The content of IMF is directly associated with the taste and flavor of pork. As a complex trait, there could be multiple genes affecting IMF content in pork. Genomewide association study is a powerful tool to detect genomic regions associated with phenotypic variations. The objectives of the present study were to identify or refine the positions of genomic regions affecting IMF, and to characterize candidate genes and pathways that may influence this trait. Of note, we identified a significant region in longissium dorsi muscle in a Duroc pig population for IMF content with Porcine SNP60 v2 Bead Chip. This region spans 1.24 Mb on chromosome 8 and had been identified as a quantitative trait locus for IMF in Pietrain, Large White, Landrace, and Leicoma pigs. In this region, eight SNPs were significantly associated with IMF content. Three genes proximal to these significant SNPs were considered candidate genes, including ZDHHC16, LOC102162218 and PCDH7. Our results confirm several previous findings and highlight several genes that may contribute to IMF variation in Duroc pigs.展开更多
Dynamic crosstalk between the embryo and mother is crucial during implantation.Here,we comprehensively profile the single-cell transcriptome of pig peri-implantation embryos and corresponding maternal endometrium,iden...Dynamic crosstalk between the embryo and mother is crucial during implantation.Here,we comprehensively profile the single-cell transcriptome of pig peri-implantation embryos and corresponding maternal endometrium,identifying 4 different lineages in embryos and 13 cell types in the endometrium.Cell-specific gene expression characterizes 4 distinct trophectoderm subpopulations,showing development from undifferentiated trophectoderm to polar and mural trophectoderm.Dynamic expression of genes in different types of endometrial cells illustrates their molecular response to embryos during implantation.Then,we developed a novel tool,ExtraCellTalk,generating an overall dynamic map of maternal-foetal crosstalk using uterine luminal proteins as bridges.Through cross-species comparisons,we identified a conserved RBP4/STRA6 pathway in which embryonic-derived RBP4 could target the STRA6 receptor on stromal cells to regulate the interaction with other endometrial cells.These results provide insight into the maternal-foetal crosstalk during embryo implantation and represent a valuable resource for further studies to improve embryo implantation.展开更多
Somatic cell nuclear transfer(SCNT)-derived piglets have signi?cantly higher stillbirth rate and postnatal mortality rate than arti?cial insemination(AI)-generated piglets. The question whether the low survival rate o...Somatic cell nuclear transfer(SCNT)-derived piglets have signi?cantly higher stillbirth rate and postnatal mortality rate than arti?cial insemination(AI)-generated piglets. The question whether the low survival rate of SCNT piglets was related to birth weight, umbilical cord or placenta development was investigated. In this study,stillbirth rate, neonatal death rate, birth weight, umbilical cord status, placental parameters and placental gene expression patterns were compared between SCNT and AI piglets. Results showed that mortality rates at birth and during the neonatal stage of SCNT piglets were signi?-cantly higher than those of AI piglets. The incidence of abnormal umbilical cord in SCNT and SCNT-liveborn(SCNT-LB) piglets was signi?cantly higher than in AI and AI-liveborn(AI-LB) piglets. Birth weight, placental weight, placental surface area and placental ef?ciency in SCNT and SCNT-LB piglets were signi?cantly lower than those of AI and AI-LB piglets. Placental expression pro?les of imprinting, angiopoiesis and nutrient transportrelated genes were defective in SCNT-LB piglets compared with those in AI-LB piglets. Thus, the low survival rate of SCNT piglets may be associated with abnormal umbilical cord and placenta development. These characteristics may have resulted from aberrant expression of angiogenesis, nutrient transport, and imprinting-related genes in the placentas.展开更多
Enucleation is a crucial procedure for mammalian somatic cell nuclear transfer(SCNT), especially for domestic animal cloning. Oocytes of domestic animals such as pigs and cattle contain dark lipid droplets that hinder...Enucleation is a crucial procedure for mammalian somatic cell nuclear transfer(SCNT), especially for domestic animal cloning. Oocytes of domestic animals such as pigs and cattle contain dark lipid droplets that hinder localization and removal of the nucleus. Using an oocyte enucleation technique that can obtain a high enucleation rate but has minimal negative effects on the reprogramming potential of oocyte for cloning is bene?cial for enhancing the outcome of SCNT. In this study, we compared the pig cloning ef?ciency resulting from blind aspiration-based(BA-B) enucleation and spindle imaging system-assisted(SIS-A) enucleation, and compared the pig SCNT success rate associated with BA-B enucleation and blind aspiration plus post-enucleation staining-based(BAPPS-B) enucleation. SIS-A enucleation achieved a signi?cantly higher oocyte enucleation success rate and tended to obtain a higher in vivo full term development rate of SCNT embryos than BA-B enucleation. BAPPS-B enucleation also obtained signi?cantly higher in vitro as well as in vivo full term development ef?ciency of cloned porcine embryos than BA-B enucleation. These data indicate that SIS-A and BAPPS-B enucleation are better approaches for pig SCNT than BA-B enucleation.展开更多
基金supported by the Guangdong Provincial Key Area Research and Development Program[grant number 2022B0202090002]China Postdoctoral Science Foundation[grant number 2024M760977].
文摘Background Hexafluoropropylene oxide dimer acid(GenX),a substitute for per-and polyfluoroalkyl substances,has been widely detected in various environmental matrices and foods recently,attracting great attention.However,a systematic characterization of its reproductive toxicity is still missing.This study aims to explore the male reproductive toxicity caused by GenX exposure and the potential cellular and molecular regulatory mechanisms behind it.Results Normally developing mice were exposed to GenX,and testicular tissue was subsequently analyzed and validated using single-cell RNA sequencing.Our results revealed that GenX induced severe testicular damage,disrupted the balance between undifferentiated and differentiated spermatogonial stem cells,and led to strong variation in the cellular dynamics of spermatogenesis.Furthermore,GenX exposure caused global upregulation of testicular somatic cellular inflammatory responses,increased abnormal macrophage differentiation,and attenuated fibroblast adhesion,disorganizing the somatic-germline interactions.Conclusions In conclusion,this study revealed complex cellular dynamics and transcriptome changes in mouse testis after GenX exposure,providing a valuable resource for understanding its reproductive toxicity.
基金supported by a Technical Innovation of Crossbred in Swine and Breed High Fertility Lines Project(2022B0202090002)a Local Innovative and Research Teams Project of Guangdong Province(2019BT02N630)+1 种基金a Natural Science Foundation of Guangdong Province project(2018B030313011)Innovative Teams of Modern Agriculture and Industry Technology System of Guangdong Province(2022KJ26).
文摘Background Pork quality can directly affect customer purchase tendency and meat quality traits have become valu-able in modern pork production.However,genetic improvement has been slow due to high phenotyping costs.In this study,whole genome sequence(WGS)data was used to evaluate the prediction accuracy of genomic best linear unbiased prediction(GBLUP)for meat quality in large-scale crossbred commercial pigs.Results We produced WGS data(18,695,907 SNPs and 2,106,902 INDELs exceed quality control)from 1,469 sequenced Duroc×(Landrace×Yorkshire)pigs and developed a reference panel for meat quality including meat color score,marbling score,L*(lightness),a*(redness),and b*(yellowness)of genomic prediction.The prediction accuracy was defined as the Pearson correlation coefficient between adjusted phenotypes and genomic estimated breeding values in the validation population.Using different marker density panels derived from WGS data,accuracy differed substantially among meat quality traits,varied from 0.08 to 0.47.Results showed that MultiBLUP outperform GBLUP and yielded accuracy increases ranging from 17.39%to 75%.We optimized the marker density and found medium-and high-density marker panels are beneficial for the estimation of heritability for meat quality.Moreover,we conducted genotype imputation from 50K chip to WGS level in the same population and found average concord-ance rate to exceed 95%and r^(2)=0.81.Conclusions Overall,estimation of heritability for meat quality traits can benefit from the use of WGS data.This study showed the superiority of using WGS data to genetically improve pork quality in genomic prediction.
基金supported by a Key Technologies R&D Program of Guangdong Province project(2022B0202090002)a China Postdoctoral Science Foundation(Grant No.2021M701263)+1 种基金a Local Innovative and Research Teams Project of Guangdong Province(2019BT02N630)a Project of Swine Innovation Team in the Guangdong Modern Agricultural Research System(2023KJ126).
文摘Background The establishment of a robust gut microbiota in piglets during their early developmental stage holds the potential for long-term advantageous effects.However,the optimal timeframe for introducing probiotics to achieve this outcome remains uncertain.Results In the context of this investigation,we conducted a longitudinal assessment of the fecal microbiota of 63 piglets at three distinct pre-weaning time points.Simultaneously,we gathered vaginal and fecal samples from 23 sows.Employing 16S rRNA gene and metagenomic sequencing methodologies,we conducted a comprehensive analysis of the fluctuation patterns in microbial composition,functional capacity,interaction networks,and colonization resistance within the gut microbiota of piglets.As the piglets progressed in age,discernible modifications in intestinal microbial diversity,composition,and function were observed.A source-tracking analysis unveiled the pivotal role of fecal and vaginal microbiota derived from sows in populating the gut microbiota of neonatal piglets.By D21,the microbial interaction network displayed a more concise and efficient configuration,accompanied by enhanced colonization resistance relative to the other two time points.Moreover,we identified three strains of Ruminococcus sp.at D10 as potential candidates for improving piglets’weight gain during the weaning phase.Conclusions The findings of this study propose that D10 represents the most opportune juncture for the introduction of external probiotic interventions during the early stages of piglet development.This investigation augments our comprehension of the microbiota dynamics in early-life of piglets and offers valuable insights for guiding forthcoming probiotic interventions.
基金supported by the National Natural Science Foundation of China (32372874)the Biological Breeding-National Science and Technology Major Project (2023ZD0404303)。
文摘Aflatoxin B_1(AFB_1) is one of the most potent mycotoxins affecting human health and animal production.To deeply understand the hosttoxin interaction,we performed CRISPR screening and identified cystathionine β-synthase(CBS) as a critical host gene affecting AFB_1 cytotoxicity.Mechanistic studies revealed that CBS affects AFB_1-induced cell death by regulating the abundance of protein post-translational modifications(PTMs) in host cells.First,AFB_1 disrupted the transfer of S-adenosylmethionine(SAM) from the cytoplasm to the mitochondria,thereby reducing the intra-mitochondrial protein methylation level.Deficient intra-mitochondrial protein methylation impaired mitochondrial function and caused cell death.CBS knockout(KO) can enhance SAM generation and mobilization to restore intramitochondrial SAM levels by rescuing the perturbed methionine cycle after AFB_1 exposure,thereby alleviating AFB_1-induced cell death.Second,AFB_1 decreased global protein ubiquitination levels by affecting gene expression of ubiquitin-modified enzymes.CBS-KO and pharmaceutical treatment correcting gene expression of ubiquitin-modified enzymes can rescue AFB_1-induced cell death.We also investigated two PTM-regulating small molecules,SAM and PR-619,which can increase cell viability in AFB_1-exposed cells.
基金supported by the National Key R&D Program(2023YFD1300202)National Natural Science Foundation of China(Grant No.32172780)Key Laboratory of Smart Agricultural Technology in Tropical South China,National Engineering Research Center for Breeding Swine Industry,and Guangdong Engineering Technology Research Center for Agricultural Farming Internet of Things.
文摘Pig body measurement is an important evaluation criterion for breeding and production management.Automatic measurement algorithms for pig body sizes exhibit sensitivity to the point cloud posture,but non-standard pig postures may result in inaccurate joint point localization in body measurement,further affecting measurement accuracy and the commercial application of these algorithms.To address this challenge,this paper proposed a pig point cloud posture transformation method based on pig’s skeleton model to adjust non-standard postures before conducting body size measurements.The method utilized an improved L1-median skeleton model to extract the three-dimensional skeleton of the pig point cloud,capturing the skeleton joint points on the target pig’s head,body,and limbs.By binding the skeleton joint points with the local point cloud and using rotation matrices,non-standard postures were adjusted to standard ones,enabling accurate body size measurements.The experimental results demonstrated that the average relative errors between the transferred posture and the original standard posture were reduced to 0.89%in body length,0.76%in body width(front),1%in body width(back),0.89%in body height(front),1.7%in body height(back),2.03%in thoracic circumference,3.37%in abdominal circumference,and 1.89%in rump circumference.To conclude,the posture standardization transfer method can significantly reduce errors in important body size parameters such as body length,body height,and body width.The method displays a greater stability and robustness compared to existing posture normalization and regression adjustment methods,providing both guidance and insight for future research in intelligent agriculture.
基金financial support from the National Natural Science Foundation of China (31601912)the Natural Science Foundation of Guangdong Province (2016A030310447)the Applied Science and Technology Research and Development of Special Funds of Guangdong Province (2015B020231010)
文摘Intramuscular fat(IMF) is a major meat-quality trait in pigs. The content of IMF is directly associated with the taste and flavor of pork. As a complex trait, there could be multiple genes affecting IMF content in pork. Genomewide association study is a powerful tool to detect genomic regions associated with phenotypic variations. The objectives of the present study were to identify or refine the positions of genomic regions affecting IMF, and to characterize candidate genes and pathways that may influence this trait. Of note, we identified a significant region in longissium dorsi muscle in a Duroc pig population for IMF content with Porcine SNP60 v2 Bead Chip. This region spans 1.24 Mb on chromosome 8 and had been identified as a quantitative trait locus for IMF in Pietrain, Large White, Landrace, and Leicoma pigs. In this region, eight SNPs were significantly associated with IMF content. Three genes proximal to these significant SNPs were considered candidate genes, including ZDHHC16, LOC102162218 and PCDH7. Our results confirm several previous findings and highlight several genes that may contribute to IMF variation in Duroc pigs.
基金supported by the National Key Research and Development Program of China(2021YFD1301103)the National Natural Science Foundation of China(31802033)+2 种基金the Guangdong Provincial Key Area Research and Development Program(2022B0202090002)the Guangdong Basic and Applied Basic Research Foundation(2022A1515011131,2023A1515030193)the Guangdong Provincial Promotion Project on Preservation and Utilization of Local Breed of Livestock and Poultry(2018143).
文摘Dynamic crosstalk between the embryo and mother is crucial during implantation.Here,we comprehensively profile the single-cell transcriptome of pig peri-implantation embryos and corresponding maternal endometrium,identifying 4 different lineages in embryos and 13 cell types in the endometrium.Cell-specific gene expression characterizes 4 distinct trophectoderm subpopulations,showing development from undifferentiated trophectoderm to polar and mural trophectoderm.Dynamic expression of genes in different types of endometrial cells illustrates their molecular response to embryos during implantation.Then,we developed a novel tool,ExtraCellTalk,generating an overall dynamic map of maternal-foetal crosstalk using uterine luminal proteins as bridges.Through cross-species comparisons,we identified a conserved RBP4/STRA6 pathway in which embryonic-derived RBP4 could target the STRA6 receptor on stromal cells to regulate the interaction with other endometrial cells.These results provide insight into the maternal-foetal crosstalk during embryo implantation and represent a valuable resource for further studies to improve embryo implantation.
基金supported by two grants received from the Department of Science and Technology of Guangdong Province, China (2016B020233006 and 2016A020210074)
文摘Somatic cell nuclear transfer(SCNT)-derived piglets have signi?cantly higher stillbirth rate and postnatal mortality rate than arti?cial insemination(AI)-generated piglets. The question whether the low survival rate of SCNT piglets was related to birth weight, umbilical cord or placenta development was investigated. In this study,stillbirth rate, neonatal death rate, birth weight, umbilical cord status, placental parameters and placental gene expression patterns were compared between SCNT and AI piglets. Results showed that mortality rates at birth and during the neonatal stage of SCNT piglets were signi?-cantly higher than those of AI piglets. The incidence of abnormal umbilical cord in SCNT and SCNT-liveborn(SCNT-LB) piglets was signi?cantly higher than in AI and AI-liveborn(AI-LB) piglets. Birth weight, placental weight, placental surface area and placental ef?ciency in SCNT and SCNT-LB piglets were signi?cantly lower than those of AI and AI-LB piglets. Placental expression pro?les of imprinting, angiopoiesis and nutrient transportrelated genes were defective in SCNT-LB piglets compared with those in AI-LB piglets. Thus, the low survival rate of SCNT piglets may be associated with abnormal umbilical cord and placenta development. These characteristics may have resulted from aberrant expression of angiogenesis, nutrient transport, and imprinting-related genes in the placentas.
基金supported by the National Natural Science Foundation of China (31772554)the Department of Science and Technology of Guangdong Province, China (2016B020233006, 2015TX01N081 and 2016A020210074)the team project of Guangdong Agricultural Department (2017LM2148)
文摘Enucleation is a crucial procedure for mammalian somatic cell nuclear transfer(SCNT), especially for domestic animal cloning. Oocytes of domestic animals such as pigs and cattle contain dark lipid droplets that hinder localization and removal of the nucleus. Using an oocyte enucleation technique that can obtain a high enucleation rate but has minimal negative effects on the reprogramming potential of oocyte for cloning is bene?cial for enhancing the outcome of SCNT. In this study, we compared the pig cloning ef?ciency resulting from blind aspiration-based(BA-B) enucleation and spindle imaging system-assisted(SIS-A) enucleation, and compared the pig SCNT success rate associated with BA-B enucleation and blind aspiration plus post-enucleation staining-based(BAPPS-B) enucleation. SIS-A enucleation achieved a signi?cantly higher oocyte enucleation success rate and tended to obtain a higher in vivo full term development rate of SCNT embryos than BA-B enucleation. BAPPS-B enucleation also obtained signi?cantly higher in vitro as well as in vivo full term development ef?ciency of cloned porcine embryos than BA-B enucleation. These data indicate that SIS-A and BAPPS-B enucleation are better approaches for pig SCNT than BA-B enucleation.
