• Journal of Applied Biological Chemistry
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• v.64 no.1
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• pp.75-81
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• 2021

Recent gut microbiota studies have revealed the important roles of gut microbiota for our health. Increasing numbers of health functional foods have been developed every year. Development of functional food often includes ex- and in-vivo experiment to verify the beneficial effects of the functional food. To investigate effects of functional food on gut microbiota, animal models were often conducted. Beneficial effects of food can be evaluated based on how gut microbiota was shifted by food, which results in either increase in beneficial bacteria, decrease in potentially pathogenic bacteria or both. As animal experiments are generally time-consuming and laborious, we investigate how well in-vitro investigation of fecal microbiota may reflect dietary health benefits. Here, we tested 15 kinds of diets using two human subjects' fecal materials. Our results showed varying gut microbiota shifts according to diets, which suggested generally known beneficial diets (i.e. Kimchi, Chunggukjang) increased Lactobacillus and Bifidobacterium. Therefore, we suggest that in vitro fecal microbiota analysis could be used to evaluate beneficial effects of diets. Moreover, this method may be ideal to establish personalized diet.

• Korean Journal of Organic Agriculture
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• v.20 no.2
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• pp.221-230
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• 2012

Rumen fermentative characteristic is useful indicators of the quality of ruminant feed stuffs and diets. An in vitro rumen fermentation experiment was therefore carried out to compare fermentation patterns among three forage sources. These were whole crop barley (WCBS), Italian ryegrass silage (IRGS) and rice straw silages (RSS). Rice straw (RS) was the control, making the treatments 4 in total. Forages were randomly allocated to serum bottles. The incubation times were arranged 0, 3, 6, 9, 12, 24, 48 and 72h at $39^{\circ}C$, respectively. Each forage source was replicated 3 times per incubation time. At each sampling time, total gas and pH were measured, whilst individual volatile fatty acids (VFAs), total volatile fatty acids (TVFAs) and ammonia nitrogen ($NH_3$-N) were determined later after storing samples at $-20^{\circ}C$. Acetate: Propionate ratio (A/P) was then calculated. Forage source had a significant effect (P<0.001) on pH and $NH_3$-N. RSS maintained higher pH values than the rest of the forage sources. A decreasing pH trend with increased time of incubation, in agreement with literature, was observed for all forage sources. WCBS recorded $NH_3$-N values higher than all the other treatments. Total gas, individual and total VFA and A/P ratio were not affected by forage source. However, there was a significant difference in all parameters (p<0.05) among forage sources at sampling periods at 3 to 72h. Therefore, the present results indicating that WCBS, IRGS, RS and RSS maintained in vitro rumen pH above the critical value. Also, WCBS produced the highest NH3-N and on this merit could be of better nutritive value, in vivo, in the ruminant.

• Animal Bioscience
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• v.35 no.12
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• pp.1892-1903
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• 2022

Objective: A series of experiment were conducted to evaluate the effects of replacing a part of soybean meal (SBM) at 6% of broiler diets with fermented soybean meal (FSBM) obtained by single or two-stage fermentation by measuring growth performance, antioxidant activity in the jejunum and distal intestinal microflora. Methods: Soybean meal samples were prepared by single-stage fermentation using Bacillus velezensis (Bv) (FSBM_B), or Lactobacillus spp. (as commercial control) (FSBM_L). Additional SBM sample was prepared by two-stage fermentation using Bv and subsequently using Lactobacillus brevis ATCC 367 (Lb) (FSBM_B+L). Enzyme activity, chemical composition, trichloroethanoic acid-nitrogen solubility index (TCA-NSI) and antioxidant activity were measured. Then, in an in vivo study, 320 Ross308 broilers were divided into four groups with ad libitum supply of feed and water. Four groups were fed either a corn-soybean meal diet (SBM), or one of fermented SBM diets (FSBM_B+L, FSBM_B, and FSBM_L). Growth, serum characteristics, microflora, and the mRNA expression of selected genes were measured. Results: Compared to SBM, FSBM_B+L contained lower galacto-oligosaccharide, allergic protein, and trypsin inhibitor, and higher TCA-NSI by about three times (p<0.05). Reducing power and 1,1-diphenyl-2-picrylhydrazyl free radical scavenging ability correlated positively with the TCA-NSI content in FSBM. Growth performances were not significantly different among four groups. In jejunum of 35-day-old broilers, partial replacement of SBM by FSBM_B+L increased the activity of superoxide dismutase and catalase (CAT), and the FSBM_B group had the highest catalase activity (p<0.05). Partial replacement of SBM by FSBM increased relative mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and peptide transporter 1 (PepT1) (p<0.05); however, FSBM_B+L increased CAT mRNA level to 5 times of the control (p<0.05). Conclusion: Using Bv- and Lb-processed SBM through two-stage fermentation to partially replace 6% of diets will improve the gut's antioxidant activity under commercial breeding in broilers.

• Nutrition Research and Practice
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• v.17 no.2
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• pp.371-385
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• 2023

BACKGROUND/OBJECTIVES: Soy isoflavone (SIF) and soy lecithin (SL) have beneficial effects on many chronic diseases, including neurodegenerative diseases. Regretfully, there is little evidence to show the combined effects of these soy extractives on the impairment of cognition and abnormal cerebral blood flow (CBF). This study examined the optimal combination dose of SIF + SL to provide evidence for improving CBF and protecting cerebrovascular endothelial cells. MATERIALS/METHODS: In vivo study, SIF50 + SL40, SIF50 + SL80 and SIF50 + SL160 groups were obtained. Morris water maze, laser speckle contrast imaging (LSCI), and hematoxylin-eosin staining were used to detect learning and memory impairment, CBF, and damage to the cerebrovascular tissue in rat. The 8-hydroxy-2'-deoxyguanosine (8-OHdG) and the oxidized glutathione (GSSG) were detected. The anti-oxidative damage index of superoxide dismutase (SOD) and glutathione (GSH) in the serum of an animal model was also tested. In vitro study, an immortalized mouse brain endothelial cell line (bEND.3 cells) was used to confirm the cerebrovascular endothelial cell protection of SIF + SL. In this study, 50 µM of Gen were used, while the 25, 50, or 100 µM of SL for different incubation times were selected first. The intracellular levels of 8-OHdG, SOD, GSH, and GSSG were also detected in the cells. RESULTS: In vivo study, SIF + SL could increase the target crossing times significantly and shorten the total swimming distance of rats. The CBF in the rats of the SIF50 + SL40 group and SIF50 + SL160 group was enhanced. Pathological changes, such as attenuation of the endothelium in cerebral vessels were much less in the SIF50 + SL40 group and SIF50 + SL160 group. The 8-OHdG was reduced in the SIF50 + SL40 group. The GSSG showed a significant decrease in all SIF + SL pretreatment groups, but the GSH showed an opposite result. SOD was upregulated by SIF + SL pretreatment. Different combinations of Genistein (Gen)+SL, the secondary proof of health benefits found in vivo study, showed they have effective anti-oxidation and less side reaction on protecting cerebrovascular endothelial cell. SIF50 + SL40 in rats experiment and Gen50 + SL25 in cell test were the optimum joint doses on alleviating cognitive impairment and regulating CBF through protecting cerebrovascular tissue by its antioxidant activity. CONCLUSIONS: SIF+SL could significantly prevent cognitive defect induced by β-Amyloid through regulating CBF. This kind of effect might be attributed to its antioxidant activity on protecting cerebral vessels.

• BMB Reports
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• v.44 no.7
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• pp.490-495
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• 2011

Transcription factor AP-$2{\alpha}$ involves in the process of mammalian embryonic development and tumorigenesis. Many studies have shown that AP-$2{\alpha}$ functions in association with other interacting proteins. In a two-hybrid screening, the regulatory subunit ${\beta}$ of protein casein kinase 2 ($CK2{\beta}$) was identified as an interacting protein of AP-$2{\alpha}$; we confirmed this interaction using in-vitro GST pull-down and in-vivo co-immunoprecipitation assays; in an endogenous co-immunoprecipitation experiment, we further found the catalytic subunit ${\alpha}$ of protein casein kinase 2 ($CK2{\alpha}$) also exists in the complex. Phosphorylation analysis revealed that AP-$2{\alpha}$ was phosphorylated by CK2 kinase majorly at the site of Ser429, and such phosphorylation could be blocked by CK2 specific inhibitor 4,5,6,7-tetrabromobenzotriazole (TBB) in a dose-dependent manner. Luciferase assays demonstrated that both $CK2{\alpha}$ and $CK2{\beta}$ enhanced the transcription activity of AP-$2{\alpha}$; moreover, $CK2{\beta}$ increased the stability of AP-$2{\alpha}$. Our data suggest a novel cellular function of CK-2 as a transcriptional co-activator of AP-$2{\alpha}$.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.12
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• pp.1798-1807
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• 2002

This study was conducted to investigate lactobacillus salivarius subsp. salivarius having probiotic properties to be used as the health adjuncts with fermented milk products. Acid- and bile-tolerant lactobacillus salivarius subsp. salivarius was isolated with lactobacilli MRS broth from faeces of 80 healthy persons (infants, children and adults). It was used as a probiotic strain in fermented milk products. The pH of fermented milk decreased from pH 6.7 to 5.0 and titratable acidity increased from 0.3% to 1.0% by L. salivarius subsp. salivarius (isolation strain 20, 35, and 37), when incubated for 36 h at 37$^{\circ}C$. The number of viable cell counts of fermented milk was maximized at this incubation condition. The SDS-PAGE evidenced no significant change of casein but distinct changes of whey protein were observed by isolated L. salivarius subsp. salivarius for titratable acidity being incubated by 0.9-1.0% at 37$^{\circ}C$. All of the strains produced 83.43 to 131.96 mM of lactic acid and 5.39 to 26.85 mM of isobutyric acid in fermented products. The in vitro culture experiment was performed to evaluate ability to reduce cholesterol levels and antimicrobial activity in the growth medium. The selected L. salivarius subsp. salivarius reduced 23-38% of cholesterol content in lactobacilli MRS broth during bacterial growth for 24 h at 37$^{\circ}C$. All of the isolated L. salivarius subsp. salivarius had an excellent antibacterial activity with 15-25 mm of inhibition zone to E. coli KCTC1039, S. enteritidis KCCM3313, S. typhimurium M-15, and S. typhimurium KCCM40253 when its pH had not been adjusted. Also, all of the isolated L. salivarius subsp. salivarius had partial inhibition zone to E. coli KCTC1039, E. coli KCTC0115 and S. enteritidis KCCM3313 when it had been adjusted to pH 5.7. The selected strains were determined to have resistances of twelve antibiotic. Strains 27 and 35 among the L. salivarius subsp. salivarius showed the highest resistance to the antibiotics. These results indicated that some of the L. salivarius subsp. salivarius (strain 27 and 35) are considered as effective probiotic strains with a potential for industrial applications, but the further study is needed to establish their use as probiotics in vivo.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.233-238
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• 2011

This study was performed to investigate the FSH levels for superovulation procedure in Korean Native Cattle (Hanwoo). The effectiveness of 200 mg and 400 mg of FSH to initiate superovulation was examined in Hanwoo. Donors, at random stages of the estrous cycle, received a CIDR 7 days later, 200 mg FSH group was treated with 40, 30, 20, 10 mg FSH levels in declining doses twice daily by intramuscular injection fur 4 days. Also, 400 mg FSH group was treated with 80, 60, 40, 20 mg FSH levels. On the 3rd day administration of FSH, 25 mg $PGF_2$ ${\alpha}$ was administered and CIDR was withdrawn. Donors were artificially inseminated twice at 12 hr intervals. The donor cattle received 250 ${\mu}g$ GnRH at time of 1st insemination and embryos were recovered 8 days after the 1st insemination. As a results, average number of CL treated with FSH 200 mg was higher as $20.9{\pm}1.20$ than $15.8{\pm}0.63$ for donors treated with FSH 400 mg, respectively(p<0.05). Treated group of 200 mg FSH level increased (p<0.05) the number of embryos recovered per procedure compared to 400 mg FSH level ($18.2{\pm}1.18$ vs. $12.38{\pm}0.52$, respectively). When treatment of 200 mg FSH was performed, average transferable embryos/ova increased (p<0.05) to $14.1{\pm}1.12$ from $6.8{\pm}0.33$ of treated of 400 mg FSH. Group of 200 mg FSH increased (p<0.05) to $8.3{\pm}0.76$ from $2.0{\pm}0.26$ in morula stage compare to 400 mg FSH group. Mean of total early blastocyst and expanded blastocyst stage embryos was similar (p<0.05) between the 200 mg and 400 mg FSH levels group ($4.7{\pm}1.19$ vs. $2.9{\pm}0.18$ and $1.2{\pm}0.40$ vs. $1.9{\pm}0.17$). These results suggest that 200 mg FSH level-based superovulation protocol with CIDR may be effectively used fur production of superior embryos in Hanwoo. In other words, the less level of FSH may be effectively applied for Hanwoo (Korean Native Cattle), because Hanwoo was smaller body size than beef or daily cow.

• Korean Journal of Veterinary Research
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• v.50 no.3
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• pp.213-220
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• 2010

Salmonella (S.) Enterica infection ranks among the most common food borne bacterial infections worldwide. Although there are six subspecies of S. Enterica, the vast majority of human and animal infections are caused by strains belonging to subspecies 1 serovar Typhimurium and Enteritidis. Recent reports on antibiotic resistance of Salmonella spp. are rising steadily. The increasing problem of antibiotic resistance has rekindled interest in bacteriophage to therapy. Therefore, we investigated the efficacy of bacteriophage in S. enterica serovar Enteritidis infected mice and pigs by measuring of body condition, body weight, bacterial colonization and weight of organs based on the in vitro analysis. In vitro experiment, phage cultured with S. Enteritidis showed clear lysis pattern, the plaque forming unit (PFU) of our phage culture was $1.5{\times}10^{11}PFU/mL$, and phage showed its maximum activity at 4 h post inoculation. In mouse experiment, there was no significant difference among experimental groups in the general body conditions and body weight of mice. However, there was difference in weight of liver and spleen depending on the experimental group (p < 0.05). The weight of liver and spleen were reduced by the phage treatment. Also bacterial colonization in spleen and liver were significantly reduced by the phage treatment. In pig experiment, the general body conditions and body temperature exhibited not much difference among the pigs except few pigs in group 3 which showed poor body conditions. From the feces in each group, we could isolate the S. Enteritidis only from group 3. Bacterial enrichment culture was necessary for isolating the bacteria from 5 dpi and 10 dpi, however direct isolation was possible from 15 dpi feces. In phage treated group, postmortem lesion was better than non-phage treated group. Recently, antibiotic resistance concerns on the food-borne bacterial pathogens have been increasing because of the wide spread of the antibiotics resistance genes. This concern is widely transmitted to the human related public health. As one of the alternative treatments on the bacterial pathogens, attempt using phages have been made to control the bacterial diseases. The positive possibility of the trail using phage was observed to control the S. enterica serovar Enteritidis in this study even though the further analysis has been remained.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.12
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• pp.1690-1695
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• 2001

Wheat straw was treated on laboratory scale with 4% urea at a moisture level of 50% along with different amounts of acetic acid (AA) to fix various levels of ammonia nitrogen (15, 30, 45 and 60%) evolved from urea and stored for 4 weeks. Chemical composition of the treated samples revealed a significant (p<0.01) increase in N content of the samples where AA was added. The N content of the ammoniated straw was only 1.21% which increased to 2.58 with the addition of AA to trap 30% $NH_3-N$. The concentration of free $NH_3-N$ in the straw was significantly (p<0.01) less when more than 15% $NH_3-N$ was trapped with AA. There was significant increase (p<0.01) in N disappearance and depression in NDF and hemicellulose disappearance, when AA was used to trap 30% $NH_3-N$. Large scale treatment of wheat straw with 4% urea at a moisture level of 50% along with AA (to trap 30% $NH_3-N$) increased the N content, but not as much as in laboratory scale treatment. In vivo experiment conducted on nine adult male buffaloes divided into three groups revealed no difference in the intake of DM, OM, NDF, ADF, cellulose and hemicellulose among group I (ammoniated straw), group II (AA treated ammoniated straw) and group III (AA treated ammoniated straw +1 kg barley grain), but the intake was significantly (p<0.05) more in groups where AA treated straw was fed as compared to only ammoniated straw fed group. However EE digestibility was depressed in group II. The digestibility of cellulose and hemicellulose both depressed significantly (p<0.05) in group II and III as compared to group I. Animals in all the 3 groups showed positive nitrogen balance and it was significantly more in group II and III as compared to group I. DCP intake was significantly (p<0.05) more in group II and III as compared to group I, but there was no significant difference among the three groups in TDN intake. It can be concluded that AA (to trap 30% $NH_3-N$) is effective in capturing the excess ammonia released during urea ammoniation of straw and improving its nutritive value, as well as animal performance.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.9
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• pp.1252-1261
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• 2008

Twenty-four yearling Boer$\times$Spanish goats were used in a crossover design experiment to determine effects of tethering on forage selection, intake and digestibility, grazing behavior and energy expenditure (EE) with forage high in nutritive value and low to moderate in mass. Objectives were to determine if tethered goats could be used as a model for study of unrestrained animals and to characterize tethering as a production practice. Four 0.72-ha pastures of wheat (Triticum aestivum) and berseem clover (Trifolium alexandrium) were grazed in December and January. Each pasture hosted six animals, three with free movement and three attached to a 4.11-m tether for access to a circular area of $53.1m^2$. Tethering areas were moved each day. One animal of each treatment and pasture was used to determine forage selection, fecal output or grazing behavior and EE; therefore, there were eight observations per treatment. Mass of forage DM before grazing in Tethered areas averaged 1,280 and 1,130 kg/ha in periods 1 and 2, respectively. The CP concentration in ingesta was greater ((p<0.05) 239 and 209 g/kg; SE = 8.0) and the NDF level was lower (p<0.05) for Free vs. Tethered animals (503 and 538 g/kg; SE = 12.0); in vitro true DM digestion was similar between treatments (0.808 and 0.807 for Free and Tethered, respectively; SE = 0.0096). Intakes of DM (1,013 and 968 g/d; SE = 78.6), NDF (511 and 521 g/d; SE = 39.9) and ME (10.9 and 10.7 MJ/d; SE = 0.90) were similar between treatments, but CP intake was greater (p<0.05) for Free vs. Tethered animals (241 and 203 g/d; SE = 17.2). There were small treatment differences in in vivo apparent digestibility of OM ((p<0.05) 0.780 and 0.814; SE = 0.0049), CP ((p<0.05) 0.800 and 0.817; SE = 0.0067) and NDF ((p<0.09) 0.777 and 0.760 for Free and Tethered, respectively; SE = 0.0078). There were no treatment effects on time spent ruminating or grazing (346 and 347 min/d for Free and Tethered, respectively; SE = 42.5), but EE was considerably greater (p<0.05) for Free vs. Tethered animals (571 and 489 kJ/kg $BW^{0.75}$; SE = 8.9). In conclusion, with forage of high nutritive value and low to moderate in mass, tethering can offer a production advantage over free grazing of less energy used for activity despite similar grazing time. With forage removal considerably less than that available for grazing, effects of tethering on chemical composition of selected forage were small and less than needed to markedly affect digestion. Tethering may offer a means of studying some aspects of grazing by ruminants, but would not seem suitable for energy metabolism.