• Journal of Animal Reproduction and Biotechnology
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• v.38 no.1
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• pp.26-31
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• 2023

Chicken embryonic stem (ES) cells have great potential and provide a powerful tool to investigate embryonic development and to manipulate genetic modification in a genome. However, very limited studies are available on the functional characterization and robust expansion of chicken ES cells compared to other species. Here, we have developed a method to generate chicken embryonic stem cell-like cells under pluripotent culture conditions. The chicken embryonic stem cell-like cells were cultivated long-term over several passages of culture without loss of pluripotency in vitro and had the specific expression of key stem cell markers. Furthermore, they showed severe changes in morphology and a significant reduction in pluripotent genes after siRNA-mediated NANOG knockdown. Collectively, these results demonstrate the efficient generation of chicken embryonic stem cell-like cells from EGK stage X blastoderm-derived singularized cells and will facilitate their potential use for various purposes, such as biobanking genetic materials and understanding stemness in the fields of animal biotechnology.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.12
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• pp.1897-1906
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• 2019

Objective: An experiment was conducted to investigate the effects of dietary non-phytate phosphorus (nPP) deficiency on intestinal pH value, digestive enzyme activity, morphology, nutrient utilization, and gene expression of NaPi-IIb in meat ducks from 1 to 21 d of age. Methods: A total of 525 one-d-old Cherry Valley ducklings were fed diets (with 7 pens of 15 ducklings, or 105 total ducklings, on each diet) with five levels of nPP (0.22%, 0.34%, 0.40%, 0.46%, or 0.58%) for 21 d in a completely randomized design. Five experimental diets contained a constant calcium (Ca) content of approximately 0.9%. Body weight (BW), body weight gain (BWG), feed intake (FI), and feed to gain ratio (F:G) were measured at 14 and 21 d of age. Ducks were sampled for duodenum and jejunum digestion and absorption function on 14 and 21 d. Nutrient utilization was assessed using 25- to 27-d-old ducks. Results: The results showed ducks fed 0.22% nPP had lower (p<0.05) growth performance and nutrient utilization and higher (p<0.05) serum Ca content and alkaline phosphatase (ALP) activity. When dietary nPP levels were increased, BW (d 14 and 21), BWG and FI (all intervals), and the serum phosphorus (P) content linearly and quadratically increased (p<0.05); and the jejunal pH value (d 14), duodenal muscle layer thickness (d 14), excreta dry matter, crude protein, energy, Ca and total P utilization linearly increased (p<0.05); however, the serum ALP activity, jejunal $Na^+-K^+$-ATPase activity, and duodenal NaPi-IIb mRNA level (d 21) linearly decreased (p<0.05). Conclusion: The results indicated that ducks aged from 1 to 21 d fed diets with 0.22% nPP had poor growth performance related to poor intestinal digestion and absorption ability; but when fed diets with 0.40%, 0.46%, and 0.58% nPP, ducks presented a better growth performance, intestinal digestion and absorption function.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.3
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• pp.403-409
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• 2018

Objective: The present study was conducted to investigate the effects of a lipid-coated zinc oxide (ZnO) supplement Shield Zn (SZ) at the sub-pharmacological concentration on intestinal morphology and gene expression in weanling pigs, with an aim to gain insights into the mechanism of actions for SZ. Methods: Forty 22-day-old weanling pigs were fed a nursery diet supplemented with 100 or 2,500 mg Zn/kg with uncoated ZnO (negative control [NC] or positive control [PC], respectively), 100, 200, or 400 mg Zn/kg with SZ for 14 days and their intestinal tissues were taken for histological and molecular biological examinations. The villus height (VH) and crypt depth (CD) of the intestinal mucosa were measured microscopically following preparation of the tissue specimen; expression of the genes associated with growth and immune function was determined using the real-time quantitative polymerase chain reaction. Results: There was no difference in daily gain, gain:feed, and diarrhea score between the SZ group and either of NC and PC. The VH and VH:CD ratio were less for the SZ group vs NC in the jejunum and duodenum, respectively (p<0.05). The jejunal mucosal mRNA levels of insulin-like growth factor (IGF-I) and interleukin (IL)-10 regressed and tended to regress (p = 0.053) on the SZ concentration with a positive coefficient, respectively, whereas the IL-6 mRNA level regressed on the SZ concentration with a negative coefficient. The mRNA levels of IGF-I, zonula occludens protein-1, tumor necrosis $factor-{\alpha}$, IL-6, and IL-10 did not differ between the SZ group and either of NC and PC; the occludin and transforming growth $factor-{\beta}1$ mRNA levels were lower for the SZ group than for PC. Conclusion: The present results are interpreted to suggest that dietary ZnO provided by SZ may play a role in intestinal mucosal growth and immune function by modulating the expression of IGF-I, IL-6, and IL-10 genes.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.12
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• pp.1742-1750
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• 2015

As a novel approach for disease control and prevention, nutritional modulation of the intestinal health has been proved. However, It is still unknown whether branched-chain amino acid (BCAA) is needed to maintain intestinal immune-related function. The objective of this study was to determine whether BCAA supplementation in protein restricted diet affects growth performance, intestinal barrier function and modulates post-weaning gut disorders. One hundred and eight weaned piglets ($7.96{\pm}0.26kg$) were randomly fed one of the three diets including a control diet (21% crude protein [CP], CON), a protein restricted diet (17% CP, PR) and a BCAA diet (BCAA supplementation in the PR diet) for 14 d. The growth performance, plasma amino acid concentrations, small intestinal morphology and intestinal immunoglobulins were tested. First, average daily gain (ADG) (p<0.05) and average daily feed intake (ADFI) (p<0.05) of weaned pigs in PR group were lower, while gain:feed ratio was lower than the CON group (p<0.05). Compared with PR group, BCAA group improved ADG (p<0.05), ADFI (p<0.05) and feed:gain ratio (p<0.05) of piglets. The growth performance data between CON and BCAA groups was not different (p>0.05). The PR and BCAA treatments had a higher (p<0.05) plasma concentration of methionine and threonine than the CON treatment. The level of some essential and functional amino acids (such as arginine, phenylalanine, histidine, glutamine etc.) in plasma of the PR group was lower (p<0.05) than that of the CON group. Compared with CON group, BCAA supplementation significantly increased BCAA concentrations (p<0.01) and decreased urea concentration (p<0.01) in pig plasma indicating that the efficiency of dietary nitrogen utilization was increased. Compared with CON group, the small intestine of piglets fed PR diet showed villous atrophy, increasing of intra-epithelial lymphocytes (IELs) number (p<0.05) and declining of the immunoglobulin concentration, including jejunal immunoglobulin A (IgA) (p = 0.04), secreted IgA (sIgA) (p = 0.03) and immunoglobulin M (p = 0.08), and ileal IgA (p = 0.01) and immunoglobulin G (p = 0.08). The BCAA supplementation increased villous height in the duodenum (p<0.01), reversed the trend of an increasing IELs number. Notably, BCAA supplementation increased levels of jejunal and ileal immunoglobulin mentioned above. In conclusion, BCAA supplementation to protein restricted diet improved intestinal immune defense function by protecting villous morphology and by increasing levels of intestinal immunoglobulins in weaned piglets. Our finding has the important implication that BCAA may be used to reduce the negative effects of a protein restricted diet on growth performance and intestinal immunity in weaned piglets.

• Journal of Animal Science and Technology
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• v.63 no.6
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• pp.1328-1343
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• 2021

The effect of Multi-Carbohydrase (MC) supplementation on growth performance, visceral organ weights, blood metabolites, jejunum morphology, nutrient digestibility, and carcass parameters of broiler chickens fed nutrient-deficient corn soybean-meal based diets containing high levels of non-starch polysaccharides from wheat and wheat by-products was investigated. A total of 378 one-day-old Ross 308 broiler chickens were randomly assigned to one of seven dietary treatments to give six replicates per treatment (nine birds per pen). Dietary treatments were as follows: (1) positive control (PC; commercial standard diet); (2) negative control 1 (NC-1; PC-120 kcal/kg metabolizable energy); (3) NC-2 (PC-3% standardized ileal digestibility [SID] amino acids). The remaining four dietary treatments were formulated with the addition of MC (MC; Superzyme-CS^TM) into two negative controls along with two supplementation levels of MC (i.e., 0.025% and 0.05%, respectively). Improved body weight, average daily gain, and feed conversion ratio (p < 0.05) were observed in broiler chickens fed a reduced energy diet supplemented with MC compared to birds fed NC-1 diet from days 1-35. Additionally, birds fed a reduced energy diet with 0.05% MC showed comparable (p > 0.05) growth performance with birds fed PC for 35-day post-hatch. Furthermore, the addition of MC into reduced amino acid diets improved (p < 0.05) growth performance. Broiler chickens fed MC supplemented nutrient-deficient diets showed a greater (p < 0.05) villus height to crypt depth ratio than birds fed diets without MC on days 21 and 35. Similarly, improved (p < 0.05) nutrient digestibility was observed in birds fed reduced energy diets supplemented with MC compared to birds fed NC-1 on days 21 and 35. Our results suggest that MC supplementation into reduced energy or reduced amino acid diets containing wheat and wheat by-products has the potential to improve growth performance and nutrient digestibility while maintaining healthier gut morphology in broiler chickens from 1 to 35 days of age.

• Asian-Australasian Journal of Animal Sciences
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• v.32 no.10
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• pp.1540-1547
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• 2019

Objective: This study was conducted to evaluate the effects of tannins and cellulase on growth performance, nutrient digestibility, blood profiles, intestinal morphology, and carcass characteristics in Hu sheep. Methods: A total of 48 three-month-old meat Hu sheep ($25.05{\pm}0.9kg$) were blocked based on body weight, and randomly allotted to 4 treatments with 3 replicates of 4 sheep each. The experiment lasted for 80 d, and dietary treatments were as follows: i) CON, control diet; ii) TAN, CON+0.1% tannins; iii) CEL, CON+0.1% cellulase; iv) TAN+CEL, CON+0.1% tannins and 0.1% cellulase. Results: Compared with CON, CEL, and TAN+CEL had greater (p<0.05) final body weight (FBW) and average daily gain but lower (p<0.05) feed conversion ratio, while FBW of TAN+CEL was lower (p<0.05) than that of CEL. The apparent total tract digestibility (ATTD) of dry matter in TAN, CEL, and TAN+CEL groups were higher (p<0.05) than that in CON. CEL and TAN+CEL groups had greater (p<0.05) ATTD of crude fiber compared with TAN and CON, while TAN group had lower (p<0.05) ATTD of crude protein than other treatments. TAN, CEL, and TAN+CEL groups increased (p<0.05) serum globulin and alkaline phosphatase but decreased (p<0.05) albumin/globulin. Serum total protein was greatest for TAN+CEL, intermediate for TAN and CEL and least for CON (p<0.05). TAN+CEL group increased (p<0.05) dressing percentage compared with CON, while the backfat thickness of CEL was lower (p<0.05) than that of CON. The villus height of jejunum and ileum in CEL and TAN+CEL groups were greater (p<0.05) than that in CON, and the crypt depth and villus height: crypt depth of jejunum were increased (p<0.05) in TAN, CEL, and TAN+CEL groups. Conclusion: The addition of tannins and cellulase together promoted nutrient digestion, liver protein synthesis and intestinal development and thus improved growth performance and carcass characteristics.

• Food Science of Animal Resources
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• v.42 no.1
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• pp.186-196
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• 2022

This study investigated the effect of exposure to flutriafol based on residues in pigs. Pigs were exposed to different concentrations (0.313, 0.625, 3.125, 6.25, and 12.5 mg/kg bw/d, n=20) for 4 wk in different treatment groups. Serum biochemical analysis, residue levels, and histological analysis were conducted using the VetTest chemistry analyzer, liquid chromatography mass spectrometry, and Masson's trichrome staining, respectively. The body weight (initial and final) was not significantly different between groups. Parameters such as creatinine, blood urea nitrogen, alanine aminotransferase, and lipase levels were significantly different as compared to the control group. Flutriafol increased the residue limits in individual tissue of the pigs in a dose dependent manner. Flutriafol exposures indicated the presence of fibrosis, as confirmed from Masson's trichrome staining. These results suggest that flutriafol affects the morphology and serum levels in pigs. The dietary flutriafol levels can provide a basis for maximum residue limits and food safety for pork and related products.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.7
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• pp.987-994
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• 2013

The purpose of this study was to evaluate the effect of natural clinoptilolite (NCLI) and modified clinoptilolite (MCLI) on broiler performance, gut morphology, intestinal length and weight, and gut digestive enzyme activity. A total of 240 d-old male chicks were randomly assigned to 3 treatments, each of which comprised 8 pens of 10 chicks per pen. Birds in the control group were fed the basal diet, while those in the experimental groups were fed diets supplemented with NCLI at 2% (NCLI group), or MCLI at 2% (MCLI group), respectively, for 42 d. Compared with the control, supplementation with NCLI or MCLI had no significant (p>0.05) effects on productive parameters from d 1 to 42. Supplementation with NCLI or MCLI had no influence on the relative length and weight of small intestine at d 1 to 21. But supplementation with NCLI or MCLI significantly reduced the relative weight of duodenum. Supplementation with MCLI and NCLI was associated with greater (p<0.05) villus height in the jejunal and ileal mucosa compared with those areas in the controls from d 1 to 42. However, supplementation with NCLI and MCLI had no significant (p>0.05) influence on the crypt depth in the jejunal and ileal mucosa compared with those in the controls. The addition of either NCLI or MCLI to the diet improved the activities of total protease, and amylase in the small intestinal contents. In conclusion, supplementation with NCLI or MCLI in diets improved intestinal morphology, increased the intestinal length and weigh and gut digestive enzyme activity.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.11
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• pp.1553-1558
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• 2002

This study was carried out to obtain informations regarding the effect of N-acetyl-D-glucosamine in the LEY (lactoseegg yolk) diluent according to incubation time in 5 ml maxi-straw and the effects of freezing rate, thawing temperature and thawing time in the LEN (lactose-egg yolk and N-acetyl-D-glucosamine) diluent on acrosome morphology and motility of frozen-thawed boar sperm. The study showed that the LEN diluent was higher post-thaw NAR (normal apical ridge) acrosome than the LEY diluent for 0.5 h incubation at 37$^{\circ}C$. However, there were no differences between the LEN and LEY diluents on post-thaw sperm motility according to incubation time. The straws frozen from 5.0 cm (20$^{\circ}C$/min) to 17.0 cm (1$^{\circ}C$/min) above the liquid nitrogen surface did not show any significant differences on post-thaw sperm motility. However, the straws frozen above 5.0 cm from the liquid nitrogen surface were higher NAR acrosome than those frozen above 17.0 cm. The post-thaw percentages of motile sperm and NAR acrosome were significantly higher (p<0.05) for the maxi-straws submerged for 40 or 45 sec in a 52$^{\circ}C$ water bath than for 30, 35, 50 or 55 sec. The mean sample temperatures of maxi-straws after 40 or 45 sec submersion were 20.7 or 26.4$^{\circ}C$. In conclusion, the sample temperature of the thawed semen was very important for post-thaw sperm survival in the LEN diluent of 5 ml maxi-straw. When the temperature of the thawed semen was 20.7$^{\circ}C$, the percentages of motile sperm and NAR acrosome were highest.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.12
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• pp.1712-1716
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• 2004

Copper-bearing montmorillonite (Cu-MMT) was produced by $Cu^{2+}$ cation exchange reaction. X-ray diffraction analysis showed that that the (001) basal spacing of the MMT crystal lattice increased from 1.544 to 1.588 nm after $Cu^{2+}$ exchange. This indicated that $Cu^{2+}$ entered into interlayer position of MMT as a hydrated cation or composite cation. In vitro results indicated that Cu-MMT had antibacterial activity on Escherichia coli $K_{88}$. Cu-MMT had unbalanced positive charge after cation exchange. Its antibacterial activity resulted from two aspects, one was electrostatic attraction which made E. coli $K_{88}$ being adhered on the montmorillonite surface, the other was the $Cu^{2+}$ slowly released, which could kill bacteria. In an in vivo study, four replicates of eight weanling pigs were assigned to each of two dietary treatments to study the effects of Cu-MMT on diarrhea, E. coli in the lumen of the jejunum and morphology of jejunal mucosa. As compared to the control, supplementation of the diet with 0.2% Cu-MMT improved average daily gain by 12.50% (p<0.05) and decreased F/G by 9.42% (p<0.05). The mean diarrhrea incidence was decreased by 71.80% (p<0.05). The viable counts of Escherichia coli in jejunal contents were significantly reduced (p<0.05). Villus height and the villus height to crypt depth ratio at the jejunal mucosa were increased by 19.09% (p<0.05) and 37.10% (p<0.05), respectively.