The full-length cDNA of the porcine peroxisomal ${\Delta}^3$,${\Delta}^2$-enoyl-CoA isomerase (PECI) gene encodes a monofunctional peroxisomal ${\Delta}^3$,${\Delta}^2$-enoyl-CoA isomerase. Cloning and sequencing of the porcine PECI cDNA revealed the presence of an 1185-base pair open reading frame predicted to encode a 394-amino acid protein by the 5'rapid amplification of cDNA ends (5'RACE) and EST sequences. The porcine PECI gene was expressed in seven tissues (heart, liver, spleen, lung, kidney, skeletal muscle, fat) which was revealed by reverse transcriptase-polymerase chain reaction (RT-PCR). The porcine PECI was mapped to SSC71/2 p11-13 using the somatic cell hybrid panel (SCHP) and the radiation hybrid panel (RH) (LOD score 12.84). The data showed that PECI was closely linked to marker S0383. A C/T single nucleotide polymorphism in PECI exon 10 (3'UTR) was detected as a PvuII PCR-RFLP. Association analysis in our experimental pig population showed that different genotypes of PECI gene were significantly associated with the Average Backfat thickness (ABF) (p<0.05) and Buttock backfat thickness (p<0.01).
The faulty regulation of imprinting gene lead to the abnormal development of reconstructed embryo after nuclear transfer. However, the correlation between the imprinting status of donor cell and preimplantation stage of embryo development is not yet clear. In this study, to determine this correlation, we used the porcine spermatogonial stem cell (pSSC) and fetal fibroblast (pFF) as donor cells. As the results, the isolated cells with laminin matrix selection strongly expressed the GFR ${\alpha}$-1 and PLZF genes of SSCs specific markers. The pSSCs were maintained to 12 passages and positive for the pluripotent marker including OCT4, SSEA1 and NANOG. The methylation analysis of H19 DMR of pSSCs revealed that the zinc finger protein binding sites CTCF3 of H19 DMRs displayed an androgenic imprinting pattern (92.7%). Also, to investigate the reprogramming potential of pSSCs as donor cell, we compared the development rate and methylation status of H19 gene between the reconstructed embryos from pFF and pSSC. This result showed no significant differences of the development rate between the pFFs ($11.2{\pm}0.8%$) and SSCs ($13.3{\pm}1.1%$). However, interestingly, while the CTCF3 methylation status of pFF-NT blastocyst was decreased (36.3%), and the CTCF3 methylation status of pSSC-NT blastocyst was maintained. Therefore, this result suggested that the genomic imprinting status of pSSCs is more effective than that of normal somatic cells for the normal development because the maintenance of imprinting pattern is very important in early embryo stage.
Chae, B.J.;Choi, S.C.;Cho, W.T.;Han, In K.;Sohn, K.S.
Asian-Australasian Journal of Animal Sciences
/
제13권10호
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pp.1440-1444
/
2000
Two feeding trials were conducted to evaluate the effects of inclusion levels of dietary vitamin and trace mineral (VTM) premixes on growth and nutrient digestibility in growing pigs. A total of 112 pigs ($24.82{\pm}3.22kg$) were employed for 49 days (exp. 1), and 168 pigs ($21.64{\pm}1.41kg$) for 40 days (exp. 2) in completely randomized block designs. Treatments were: 1) 100%, 2) 150%, 3) 200% and 4) 250% NRC (1998) requirement of VTM in exp. 1, and the ratio of vitamins to trace minerals at 1) 100:100%, 2) 100:150%, 3) 150:100% and 4) 150:150% of NRC (1998) requirement in exp. 2. Basal diets for feeding trials were formulated to contain 3,310 kcal ME/kg and 18% crude protein, and contained 0.25% chromic oxide as an indigestible marker for digestibility trials. Increasing dietary VTM premix in growing pigs had linear and quadratic effects (p<0.05) on ADG, and feed conversion ratio was also improved (p<0.05) as VTM premix was increased by 150-250% of NRC (1998) requirements in exp. 1. Adding vitamin to trace mineral premixes at 150% NRC (1998) over the control improved (p<0.05) ADG and feed efficiency in growing pigs, but performances were not improved by vitamin nor by trace mineral premixes alone (p>0.15) (exp. 2). There were no differences (p>0.05) in the digestibilities of energy, crude protein and fat among dietary treatments. However, increasing dietary VTM premix in growing pigs had a linear effect (p<0.05) on the digestibilities of calcium and phosphorus. The 200 or 250% fed group showed improved (p<0.05) calcium digestibility, and 250% fed group also showed improved (p<0.05) phosphorus digestibility as compared to 100% or 150% fed group (exp. 1). The digestibilities of Ca and P were higher (p<0.05) in 150% addition of vitamins than in 150% addition of trace minerals in the diet (exp. 2).
Two experiments were conducted to compare the effects of feeding a newly-developed rare earth mineral-yeast product, zinc oxide (ZnO) or antibiotics on the performance, nutrient digestibility and serum parameters of weanling pigs. In experiment 1, 150 crossbred barrows (24 d old and 6.28 kg BW) were fed one of five dietary treatments consisting of an unsupplemented basal diet or the basal diet supplemented with antibiotics (33 ppm tiamulin and 100 ppm chlortetracycline), ZnO (1,500 or 2,500 ppm) or 0.1% peptide-bound rare earth mineral-yeast. In experiment 2, 576 crossbred barrows (28 d old and 7.20 kg BW) were fed the same diets as those used in experiment 1 modified only by the addition of 1.0% Celite 545 to all diets as a digestibility marker. However, the negative control was not included. In experiment 1, weight gain was significantly lower (p<0.05) for pigs fed the negative control than for pigs fed diets supplemented with antibiotics, ZnO, or rare earth mineral-yeast. Pig performance did not differ between pigs fed the four supplemented diets. In experiment 2, there were no differences in performance between pigs fed diets supplemented with antibiotic, ZnO or rare earth mineral-yeast. The digestibility of dry matter, crude protein, calcium, phosphorus and energy were significantly (p<0.01) higher on the rare earth mineral-yeast diet than on diets supplemented with ZnO. In addition, pigs fed the diet supplemented with rare earth mineral-yeast had significantly (p<0.05) higher digestibility of histidine, lysine, threonine and valine than pigs fed the ZnO supplemented diets. Digestibility coefficients for pigs fed antibiotics tended to be intermediate to those of pigs fed rare earth mineralyeast or ZnO. In conclusion, the performance of pigs fed rare earth mineral-yeast was basically equal to that of pigs fed antibiotics or ZnO indicating that rare earth mineral-yeast can be successfully used as a growth promoter in diets fed to nursery pigs. The effects of rare earth mineral-yeast appeared to be mediated through improvements in nutrient digestibility.
The purpose of this study was to develop the diagnosis techniques for sex determination of rabbit embryos at preimplantation stage. To detect male specific sequences using polymerase chain reaction, two genes functional on sex determination including SRY and ZFX/Y genes were targeted using multiple oligonucleotide primer sets. Three of them for conserved SRY gene were used for appropriate amplification pattern, and then only one primer set #3 proved to be most efficient, showing male-specific strong signal ofamplified sequences. Using this male specific bandsfrom human, cattle, pig and mouse, the gender of rabbit was determined. As an another system for sex determination system, amplified 910bp fragment from ZFX/Y was digested with several restriction endonuclease and showed gender specific restriction fragments only by Hinf I. Using two different system for sex identification of rabbit in this study, blind tests for 17 samples was conducted and showed identical results from two different methods. And then, amplification limit of PCR reaction for template DNA was estimated using various amounts of DNA for both SRY and ZFX/Y systems, resulted as 20pg and 800pg, respectively. With this results, test for gender identification of rabbit embryos were performed using SRY derived amplification system. From total 22 embryos selected for its developmental state 18 were identified as male embryos, showing significant difference from expected sex ratio 1:1. This biased sex ratio was interpreted as to have been caused by the fact, reported by the fact, reported by several researchers, that male embryos develop more rapidly and are more resistant against the in vitro manipulation than female embryos.
Zhang, Yongcheng;Li, Defa;Fan, Shijun;Piao, Xiangshu;Wang, Jitan;Han, In K.
Asian-Australasian Journal of Animal Sciences
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제15권11호
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pp.1634-1638
/
2002
Quantification of endogenous amino acid loss at the terminal ileum is an essential means for calculation of the true amino acid digestibility of a feedstuff. Since nitrogen appeared in the determined diet or not could shift the results very much, also, none of digestibility markers could be recovered with 100% rate at the terminal ileum, the objectives of the present study were: (1) to determine endogenous amino acid losses when fed either a casein diet or a protein-free diet and (2) to examine the reliability of chromic oxide or acid insoluble ash in the protein-free diet. Six ileal-cannulated pigs ($65{\pm}1.85 kg$ BW) with a simple T-cannula in the terminal ileum were used in a replicated $3{\times}3$ Latin square designed trial, after allowed a 14 d recuperation period. Each test period ran for 12 days comprised of a 10 d adjustment period and a 2 d collection period. The endogenous AA losses of His, Ile, Lys, Cys, Thr, Val, Trp, Asp, Glu, and Ser from pigs fed the casein diet were significantly higher than those of the protein-free diet (p<0.05). No significant difference was found in the amount of endogenous amino acid loss when determined with the different markers in the protein-free diet (p>0.05). These data suggest that endogenous amino acid loss could be underestimated when a protein-free diet is used. A direct effect of dietary peptides on the endogenous amino acid loss was found when the casein diet was fed. Our results also indicate that acid insoluble ash can be used as an inert marker as an alternative to chromic oxide when measuring endogenous amino acid loss.
This study was undertaken to examine the influence of soluble non-starch polysaccharides on growth performance, mucin secretion, and endogenous amino acid flows in weaner pigs. Different levels (0, 4 and 7.5%) of purified corn arabinoxylan (AX) or barley $\beta$-glucan extract (BG) were substituted for cellulose in a purified diet based on starch, sucrose and enzymatically hydrolyzed casein. All diets contained titanium oxide as an indigestible marker. Each experimental diet was fed to five, 6-wk old weaner pigs for 21 days. Average daily gain (p<0.05) and feed conversion ratio (p<0.01) were improved with dietary inclusion of 7.5% AX and BG, indicating high degradation rates of AX and BG in pigs. Crude mucin contents and endogenous nitrogen flow were increased (p<0.05) with increased levels of AX, but not with BG. Numerical increases in endogenous amino acid flow (EAAF) were observed with increased levels of AX but no definite trend with BG. Endogenous amino acid flow in pigs fed mixed NSP diets (4% BG and 3.5% cellulose) was significantly higher (p<0.05) than those fed 7.5% BG diets. Among diets containing pure sources of soluble non-starch polysaccharides, endogenous amino acid flows were highest in 7.5% AX (p<0.05), intermediate in BG, and lowest in control diet. Increased flows (p<0.01) of threonine, proline and serine in pigs fed 7.5% AX diets are consistent with the increased flow of crude mucin determined in this treatment. In conclusion, mucin and endogenous amino acid flows were increased with dietary inclusion of AX, which could be related to its physicochemical property, particularly its high water-holding capacity. In contrast, $\beta$-glucan, due to its high degradation rate in pig, may be considered as unimportant factor in inducing mucin and endogenous amino acid secretions, at least at levels such as those used in this study.
Objective: The objective of a conservation program is to maintain maximum genetic diversity and preserve the viability of a breed. However, the efficiency of a program is influenced by the ability to accurately measure and predict genetic diversity. Methods: To examine this question, we conducted a simulation in which common measures (i.e. heterozygosity) and novel measures (identity-by-descent probabilities and parental genomic components) were used to estimate genetic diversity within a conserved population using double-labeled single nucleotide polymorphism markers. Results: The results showed that the accuracy and sensitivity of identity-by-state probabilities and heterozygosity were close to identity by descent (IBD) probabilities, which reflect the true genetic diversity. Expected heterozygosity most closely aligned with IBD. All common measures suggested that practices used in the current Chinese pig conservation program result in a ~5% loss in genetic diversity every 10 generations. Parental genomic components were also analyzed to monitor real-time changes in genomic components for each male and female ancestor. The analysis showed that ~7.5% of male families and ~30% of female families were lost every 5 generations. After 50 generations of simulated conservation, 4 male families lost ~50% of their initial genomic components, and the genomic components for 24.8% of the female families were lost entirely. Conclusion: In summary, compared with the true genetic diversity value obtained using double-labeled markers, expected heterozygosity appears to be the optimal indicator. Parental genomic components analysis provides a more detailed picture of genetic diversity and can be used to guide conservation management practices.
The purpose of this study was to establish a rapid, reliable diagnostic method detecting Encephalomyocarditis virus(EMCV) RNA in formalin-fixed, paraffin-embedded tissues of EMCV naturally infected pigs by cDNA probe of EMC $K_3$, the EMCV strain isolated from Korea. Using a biotin-labelled nick translated probe for the cDNA marker. We made up for some defects of radiolabeled method. In sits hybridization(ISH) technique, differently from the other nucleic acid hybridization methods, is able to detect the virus genome specifically in the state of the intact shapes of cells and/or tissues. We succeeded in performing the experiment to detect the EMCV within 1~2 hours using the $MicroProbe^{TM}$ capaillary action system. In this study, we observed highly specific positive signals of red color by staining the paraffin-embedded tissue sections of naturally EMCV-infected pig organs or tissues, including brain, heart, kidney and lacrimal gland with the Fast Red TR salt/Naphtol phosphate chromogen. The results suggested that this ISH method is considered as a highly sensitive and reliable tool for molecular biologic diagnosis of the EMC viral disease.
The acute phase serum protein response is a well-known general indicator of inflammation, trauma or other pathological conditions and its relevance for the monitoring of the health status of domestic animals is being increasingly realized. The changes in serum protein composition which occur after tissue damage represent a part of the systemic response of the injured animals which is mediated by pro-inflammatory cytokines such as TNF-$\alpha$, IL-6 and IL-1. These responses play a vital role in containing the tissue damage and enhancing the processes of repair and resolution. From a clinical perspective, the assay of acute phase proteins can provide a method for detecting inflammation. In animals, the most sensitive acute phase proteins are haptoglogin, serum amyloid A and at-acid glycoprotein in response to inflammatory condition. The aim of this study was to assess the diagnostic value of the concentrations of serum amyloid A(SAA) and haptoglobin(HP) in serum of pigs infected with Aujeszky's disease virus(ADV). Fifty pigs infected with ADV and 5 normal pigs were used in this experiment. The mean serum concentration of Shh of pigs infected with ADV was 96.8 $\pm$ 7.1 $\mu\textrm{g}$/㎖(range, 36.0∼187.5 $\mu\textrm{g}$/㎖) and that of normal pigs was 42.9$\pm$3.3 $\mu\textrm{g}$/㎖(range, 17.3∼127.8 $\mu\textrm{g}$/㎖). The mean serum concentration of HP of pigs infected with ADV was 1,164.4 $\pm$ 96.9 $\mu\textrm{g}$/㎖ (range, 790.2∼l,769.2 $\mu\textrm{g}$/㎖) and that of normal pigs was 675.4 $\pm$ 56.3 $\mu\textrm{g}$/㎖ (range, 650.0-690.4 $\mu\textrm{g}$/㎖). The mean concentrations of SAA and HP in serum of pigs infected with ADV compared with those of normal pigs showed approximately a two-fold. It was concluded that the concentrations of Shh and HP in serum may proved to be diagnostic marker of Aujeszky's disease.
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