Kim, Min-Goo;Seo, Hee-Won;Choi, Yo-Han;Shim, Jang-Soo;Kim, Hee-Bal;Lee, Chang-Kyu;Ka, Hak-Hyun
Asian-Australasian Journal of Animal Sciences
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v.25
no.8
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pp.1102-1116
/
2012
During embryo implantation in pigs, the uterine endometrium undergoes dramatic morphological and functional changes accompanied with dynamic gene expression. Since the greatest amount of embryonic losses occur during this period, it is essential to understand the expression and function of genes in the uterine endometrium. Although many reports have studied gene expression in the uterine endometrium during the estrous cycle and pregnancy, the pattern of global gene expression in the uterine endometrium in response to the presence of a conceptus (embryo/fetus and associated extraembryonic membranes) has not been completely determined. To better understand the expression of pregnancy-specific genes in the endometrium during the implantation period, we analyzed global gene expression in the endometrium on day (D) 12 and D15 of pregnancy and the estrous cycle using a microarray technique in order to identify differentially expressed endometrial genes between D12 of pregnancy and D12 of the estrous cycle and between D15 of pregnancy and D15 of the estrous cycle. Results showed that the global pattern of gene expression varied with pregnancy status. Among 23,937 genes analyzed, 99 and 213 up-regulated genes and 92 and 231 down-regulated genes were identified as differentially expressed genes (DEGs) in the uterine endometrium on D12 and D15 of pregnancy compared to D12 and D15 of the estrous cycle, respectively. Functional annotation clustering analysis showed that those DEGs included genes involved in immunity, steroidogenesis, cell-to-cell interaction, and tissue remodeling. These findings suggest that the implantation process regulates differential endometrial gene expression to support the establishment of pregnancy in pigs. Further analysis of the genes identified in this study will provide insight into the cellular and molecular bases of the implantation process in pigs.
Lee, Won Young;Lee, Ran;Kim, Hee Chan;Lee, Kyung Hoon;Cui, Xiang Shun;Kim, Nam Hyung;Kim, Sang Hyun;Lee, Il Joo;Uhm, Sang Jun;Yoon, Min Jung;Song, Hyuk
Asian-Australasian Journal of Animal Sciences
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v.27
no.10
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pp.1417-1425
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2014
The selection of morphologically normal spermatozoa is critical to obtain high breeding performances in boar breeding farms and artificial insemination (AI) centers. Parameters for the selection of semen mainly include total sperm motility, concentration, and morphology. However, these primary parameters are often not reliable for discriminating between normal and abnormal, non-fertilizable spermatozoa. The present study was designed to compare the motion characteristics, fertilization ability using in vitro fertilization (IVF), and acrosome formation of the semen from boars having low (boar number 2012) and normal (boar number 2004 and 2023) breeding performances. The ultimate goal was to identify additional simple and easy criteria for the selection of normal sperm. There was no significant difference between boar 2004 and boar 2023 sperm total motility in computer assisted sperm analysis. However, boar number 2012 semen presented a significantly reduced population of rapid moving spermatozoa and an increased population of slow moving spermatozoa compared to boar numbers 2004 and 2023. Analysis of detailed motion characteristics revealed that sperm from boar number 2012 had significantly reduced motility in progressiveness, average path velocity, straight-line velocity (VSL), curvilinear velocity (VCL), straightness, and linearity. The assessment of the fertilizing ability by IVF also showed that sperm from boar number 2012 showed a fertility rate of 3.4%, whereas sperm from boar number 2023 had a fertility rate of 75.45%. Interestingly, most of the sperm nuclei were found on the peripheral area of the oocytes, suggesting that the sperm from boar number 2012 lacked penetration ability into the oocyte zonapellucida. The acrosome formation analysis using Pisum sativum agglutinin staining demonstrated that the sperm from boar number 2012 had a defect in acrosome formation. Consequently, primary parameters for selecting semen before AI such as motility are not sufficient to select normal and fertilizable spermatozoa. In conclusion, the present study suggests that the acrosome staining and detailed motion characteristics such as progressiveness, VCL, and VSL should be included in determining semen quality together with primary parameters for successful AI and high breeding performance in the swine industry.
Cluster of differentiation 4 (CD4) is mainly expressed on $CD4^+$ T cells, which plays an important role in immune response. The aim of this study was to detect the association between polymorphisms of the CD4 gene and T lymphocyte subpopulations in pigs, and to investigate the effects of genetic variation on the CD4 gene expression level in immune tissues. Five missense mutations in the CD4 gene were identified using DNA pooling sequencing assays, and two main haplotypes (CCTCC and AGCTG) in strong linkage disequilibrium (with frequencies of 50.26% and 46.34%, respectively) were detected in the population of Large White pigs. Our results indicated that the five SNPs and the two haplotypes were significantly associated with the proportions of $CD4^-CD8^-$, $CD4^+CD8^+$, $CD4^+CD8^-$, $CD4^+$ and $CD4^+/CD8^+$ in peripheral blood (p<0.05). Gene expression analysis showed the mRNA level of the CD4 gene in thymus was significantly higher than that in lymph node and spleen (p<0.05). However, no significant difference was observed between animals with CCTCC/CCTCC genotype and animals with AGCTG/AGCTG genotype in the three immune tissues (p>0.05). These results indicate that the CD4 gene may influence T lymphocyte subpopulations and can be considered as a candidate gene affecting immunity in pigs.
Lv, J.N.;Chen, Y.Q.;Guo, X.J.;Piao, X.S.;Cao, Y.H.;Dong, B.
Asian-Australasian Journal of Animal Sciences
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v.26
no.4
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pp.579-587
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2013
A total of 288 crossbred (Duroc${\times}$Landrace${\times}$Yorkshire) growing pigs were used in two experiments to investigate the effects of adding ${\beta}$-mannanase to corn-soybean meal-based diets on pig performance and apparent total tract digestibility (ATTD). Both experiments lasted 28 d and were split into two phases namely 1 to 14 days (phase 1) and 15 to 28 days (phase 2). In Exp. 1,144 pigs weighing $23.60{\pm}1.59$ kg BW were assigned to one of four corn-soybean meal-based diets containing 0, 200, 400 or 600 U/kg ${\beta}$-mannanase. Increasing the level of ${\beta}$-mannanase increased weight gain (quadratic effect; p<0.01) and feed efficiency (linear and quadratic effect; p<0.01) during the second phase and the overall experiment. However, performance was unaffected (p>0.05) by treatment during phase 1. Increasing the amount of ${\beta}$-mannanase in the diet improved (linear and quadratic effect; p<0.05) the ATTD of CP, NDF, ADF, calcium, and phosphorus during both phases. Based on the results of Exp. 1, the optimal supplementation level was determined to be 400 U/kg and this was the level that was applied in Exp. 2. In Exp. 2, 144 pigs weighing $23.50{\pm}1.86$ kg BW were fed diets containing 0 or 400 U/kg of ${\beta}$-mannanase and 3,250 or 3,400 kcal/kg digestible energy (DE) in a $2{\times}2$ factorial design. ${\beta}$-Mannanase supplementation increased (p<0.01) weight gain and feed efficiency while the higher energy content increased (p<0.01) feed intake and feed efficiency during both phases and overall. Increased energy content and ${\beta}$-mannanase supplementation both increased (p<0.05) the ATTD of DM, CP, NDF, ADF, phosphorus, and GE during both phases. There were no significant interactions between energy level and ${\beta}$-mannanase for any performance or digestibility parameter. In conclusion, the ${\beta}$-mannanase used in the present experiment improved the performance of growing pigs fed diets based on corn and soybean. The mechanism through which the improvements were obtained appears to be related to improvements in ATTD.
Forty-eight barrows with an average initial body weight of $25.5{\pm}0.3kg$ were assigned to 6 dietary treatments arranged in a $3{\times}2$ factorial of 3 graded levels of P at 1.42, 2.07, or 2.72 g/kg, and 2 levels of casein at 0 or 50 g/kg to compare the estimates of true total tract digestibility (TTTD) of P in soybean meal (SBM) for pigs fed diets with or without casein supplementation. The SBM is the only source of P in diets without casein, and in the diet with added casein, 1.0 to 2.4 g/kg of total dietary P was supplied by SBM as dietary level of SBM increased. The experiment consisted of a 5-d adjustment period and a 5-d total collection period with ferric oxide as a maker to indicate the initiation and termination of fecal collection. There were interactive effects of casein supplementation and total dietary P level on the apparent total tract digestibility (ATTD) and retention of P (p<0.05). Dietary P intake, fecal P output, digested P and retained P were increased linearly with graded increasing levels of SBM in diets regardless of casein addition (p<0.01). Compared with diets without casein, there was a reduction in fecal P in the casein-supplemented diets, which led to increases in digested P, retained P, ATTD, and retention of P (p<0.01). Digested N, ATTD of N, retained N, and N retention were affected by the interaction of casein supplementation and dietary P level (p<0.05). Fecal N output, urinary N output, digested N, and retained N increased linearly with graded increasing levels of SBM for each type of diet (p<0.01). The estimates of TTTD of P in SBM, derived from the regression of daily digested P against daily P intake, for pigs fed diets without casein and with casein were calculated to be 37.3% and 38.6%, respectively. Regressing daily digested N against daily N intake, the TTTD of N in SBM were determined at 94.3% and 94.4% for diets without casein and with added casein, respectively. There was no difference in determined values of TTTD of P or N in SBM for pigs fed diets with or without casein (p>0.05). In summary, our results demonstrate that the estimates of TTTD of P in SBM for pigs were not affected by constant casein inclusion in the basal diets.
Objective: Prostaglandins (PGs) function in various reproductive processes, including luteolysis, maternal pregnancy recognition, conceptus development, and parturition. Our earlier study has shown that PG transporters ATP-binding cassette, subfamily C, member 4 (ABCC4) and solute carrier organic anion transporter family, member 2A1 (SLCO2A1) are expressed in the uterine endometrium in pigs. Since several other PG transporters such as ABCC1, ABCC9, SLCO4C1, and SLCO5A1 are known to be present in the uterine endometrium, this study investigated the expression of these PG transporters in the porcine uterine endometrium and placenta. Methods: Uterine endometrial tissues were obtained from gilts on day (D) 12 and D15 of the estrous cycle and days 12, 15, 30, 60, 90, and 114 of pregnancy. Results: ABCC1, ABCC9, SLCO4C1, and SLCO5A1 mRNAs were expressed in the uterine endometrium, and levels of expression changed during the estrous cycle and pregnancy. Expression of ABCC1 and ABCC9 mRNAs was localized mainly to luminal and glandular epithelial cells in the uterine endometrium, and chorionic epithelial cells during pregnancy. Conceptuses during early pregnancy and chorioallantoic tissues from mid to late pregnancy also expressed these PG transporters. $Estradiol-17{\beta}$ increased the expression of ABCC1 and SLCO5A1, but not ABCC9 and SLCO4C1 mRNAs and increasing doses of $interleukin-1{\beta}$ induced the expression of ABCC9, SLCO4C1, and SLCO5A1 mRNAs in endometrial explant tissues. Conclusion: These data showed that several PG transporters such as ABCC1, ABCC9, SLCO4C1, and SLCO5A1 were expressed at the maternal-conceptus interface, suggesting that these PG transporters may play an important role in the establishment and maintenance of pregnancy by regulating PG transport in the uterine endometrium and placenta in pigs.
Zhao, Jinbiao;Li, Zhongchao;Lyu, Mingbin;Liu, Ling;Piao, Xiangshu;Li, Defa
Asian-Australasian Journal of Animal Sciences
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v.30
no.5
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pp.712-719
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2017
Objective: The objective of this study was to determine the effect of inclusion level on the digestible energy (DE), metabolizable energy (ME), and total tract digestibility of acid-hydrolyzed ether extract (AEE) of cottonseed oil when fed to growing pigs. Methods: Forty-two barrows (initial body weight = $35.51{\pm}2.01kg$) were randomly allotted to a completely randomized design with a corn-soybean meal basal diet, five levels of cottonseed oil (2%, 4%, 6%, 8%, and 10%) and a 10% soybean oil diet. Each diet was replicated six times with one pig per replicate. The experiment lasted 19 days, 7 d for cage adaptation, 7 d for diets adaptation and last 5 d for feces and urine collection. The energy values and apparent total tract digestibility (ATTD) of cottonseed oil and soybean oil were calculated by the difference method, and regression equations were established to predict the energy values of cottonseed oil. The apparent digested fat of the entire intestinal tract was also regressed against dietary fat intake to determine the true total tract digestibility (TTTD) and endogenous loss of fat for cottonseed oil. Results: The results showed that the DE and ME contents of cottonseed oil were not different as the inclusion level increased. The DE and ME values determined by the regression equation were 36.28 MJ/kg and 34.96 MJ/kg, respectively, and the values were similar to the mean DE and ME values calculated by the difference method (36.18 and 35.56 MJ/kg, respectively). The ATTD of cottonseed oil was also not affected by the inclusion level of cottonseed oil, and the TTTD and EFL determined by the regression method were 92.40% and 13.83 g/kg of dry matter intake for corn-soybean basal diet. The DE, ME, and ATTD of AEE in soybean oil determined by the difference method were 35.70 MJ/kg, 35.20 MJ/kg and 92.31%, respectively. There were no differences in the DE, ME, and ATTD between cottonseed oil and soybean oil, although the ratio of unsaturated to saturated fatty acids for soybean oil was higher than for cottonseed oil. Conclusion: The DE, ME, and ATTD values of cottonseed oil were not affected by its dietary inclusion level. The energy values of cottonseed oil determined by the difference and regression methods were similar. Furthermore, the ratio of unsaturated to saturated fatty acid for oils was not the decisive factor to influence the energy values and ATTD of oils.
Objective: This study was conducted to investigate the effects of dietary probiotic blend and liquid feed program at different nutritional densities on growth performance, nutrient digestibility, fecal score of weaning piglets. Methods: A total of 120 weaning pigs with an initial body weight of 7.05±0.93 kg per pig (21 days of age) were randomly allocated into 1 of the following 8 dietary treatments (3 replicates per treatment with 5 pigs per replicate) in a 2×2×2 factorial arrangement (nutrition levels: apparent metabolic energy [AME] = 3,500 kcal/kg, crude protein [CP] = 20% vs AME = 3,400 kcal/kg, CP = 19.42%; feed types:dry vs wet; probiotics levels: 0 mg/kg vs 300 mg/kg). Results: During d 5 to d 15, greater average daily gain (ADG) and average daily feed intake (ADFI) (p<0.05) were observed in probiotics treatments. During d 15 to d 25, gain:feed (G:F) ratio (p<0.05) were significantly improved in probiotics, wet feed and high nutrition diet. Moreover, two interactions i) between nutrition levels and feed types, and ii) between nutrition levels and probiotics were found in G:F ratio. Furthermore, there was a significant positive interaction on G:F among those 3 factors (p<0.05). Overall, increasing ADG, ADFI, and G:F ratio were detected in probiotics treatment significantly (p<0.05). Besides, an obvious reduction on fecal score was observed in probiotics treatment from d 0 to d 5 (p<0.05). There was an interactive effect on fecal score between feed types and nutrition concentrations from d 5 to d 25 (p<0.05). Conclusion: These results indicated that probiotics supplementation could benefit growth performance and reduce the frequency of watery feces. Besides, wet feed program (feed:water = 1:1.25) could improve the G:F. The effect of liquid feed or probiotic could be influenced by dietary nutrition density in weaned piglets. An increased value of G:F was obtained when wet feeding a high nutrition diet (100 kcal higher than NRC 2012 recommendations) was supplemented with probiotics for 15 to 25 days.
Kim, B.G.;Tian, J.Z.;Lim, J.S.;Kil, D.Y.;Jeon, H.Y.;Chung, Y.K.;Kim, Y.Y.
Asian-Australasian Journal of Animal Sciences
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v.17
no.12
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pp.1729-1735
/
2004
A total of 140 weaning pigs were used to determine the effects of digestive enzyme supplementation to corn-soybean meal diets on growth performance, physiological changes of small intestine, microorganisms and pH in the gastrointestinal tract. Two kinds of enzyme complex (A, B) were used in this experiment. Pigs were allotted in a completely random design (CRD) to five replicates with four pigs per pen. Diets and water were provided for ad libitum consumption. Treatments included 1) Control: without enzyme supplementation, 2) Enzyme A 0.05%, 3) Enzyme A 0.10%, 4) Enzyme A 0.15%, 5) Enzyme B 0.05%, 6) Enzyme B 0.10%, 7) Enzyme B 0.15% in the diets. A total of 24 crossbred barrows 25.78${\pm}$0.55 kg BW fitted with simple ileal T-cannulas were used to evaluate the effect the enzyme addition on the nutrient digstibility. Pigs were allotted 4 treatments (No enzyme, enzyme A 0.05%, enzyme A 0.1%, enzyme A 0.15%), 6 replicates according to a completely random design (CRD). Another digestibility trial was followed for enzyme complex B. Twenty pigs, average 31.92${\pm}$0.37 kg BW, fitted with simple ileal T-cannulas for digestibility trial. Neither enzyme A nor enzyme B affected on fecal or ileal digestibility of dry matter, gross energy, crude protein, crude fat and crude ash (p>0.05). The apparent fecal digestibilities of all the nutrients were higher in total feces collection method than in indirect method. At the end of feeding trial, 21 pigs were slaughtered for examining the morphological changes of small intestine and the concentration of microorganisms in the ileum and the colon. Growth performance, intestinal morphology and pH of ileum and colon were not affected by the either enzyme complex supplementation (p>0.05). These results suggested that enzyme complex A and enzyme complex B were of no benefit to early-weaned pigs when corn-soybean meal based diet was provided.
Previous studies conducted with swine have reported that the mobile nylon bag technique (MNBT) does not always accurately predict in vivo nutrient digestibilities. Therefore, in this study, the MNBT was modified so that nutrient digestibilities would more closely resemble those from conventional (Con) digestibility studies obtained using the indicator method. A total of 19 feeds were tested including five cereal grains, five legumes, three high protein sources and six mixed diets. The principle changes to the MNBT included the use of a fecal collection harness which minimized the number of bags lost. In addition, previous protocols involved pooling of bags within pig while in the present experiment all bags were analyzed separately to increase the precision of the test. Finally, chemical analyses were done using the entire nylon bag plus residue rather than opening.the bags and scraping out the contents. With the exception of the barley sample (p=0.01), dry matter digestibility (DMD) coefficients obtained with the MNBT were not significantly different from those obtained with the indicator method. The linear regression equation relating the MNBT to the indicator method was Con DMD=-O.77+1.02 MNBT DMD ($r^2=0.93$: p<0.0001). There was no significant (p>0.05) difference in gross energy digestibility (GED) coefficients determined using the MNBT or the indicator method for any of the 19 feeds. The regression line equation relating the MNBT to the indicator method was Con GED=-5.68+1.06 MNBT GED ($r^2=0.94$: p<0.0001). The MNBT was less effective in predicting in vivo crude protein digestibility (CPD) than it was in predicting dry matter and energy digestibility. Differences greater than five percentage units were observed for two of the legumes, Kabuli chickpeas (p=0.02) and the extruded pea-canola seed mixture (p=0.01) as well as for three of the mixed diets including the unheated hulled barley-based diet (p=0.01), the unheated hulless-barley based diet (p=0.08) and the barley-soybean meal based diet (p=0.008). The regression equation relating the MNBT to the indicator method was Con CPD=5.75 + 0.90 MNBT CPO ($r^2=0.76$; p<0.0001). This study indicates that the modified MNBT can be used for the rapid determination of dry matter and energy digestibility in a wide variety of ingredients. For the measurement of crude protein digestibility, the technique produces results similar to conventional digestibility studies for cereal grains and high protein feeds but tends to overestimate protein digestibility for legumes and mixed diets.
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