• Journal of Life Science
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• v.31 no.10
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• pp.922-928
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• 2021

To profile the microbial compositions of Korean traditional fermented paste made from whole soybeans, Doenjang and Cheonggukjang, and compare their taxonomic differences, we analyzed the V3-V4 region of 16S rRNA of naturally fermented foods by using next generation sequencing. α-Diversity results showed that values indicating bacterial community abundances (OTUs) and richness (ACE, Chao1) were statistically significant (p=0.0001) in Doenjang and Cheonggukjang. Firmicutes was the most common phylum in both groups, representing 97.02% and 99.67% in the Doenjang and Cheonggukjang groups, respectively. Bacillus was the most dominant genus, accounting for 71.70% and 59.87% in both groups. Linear discriminant (LDA) effect size (LEfSe) analysis was performed to reveal the significant ranking of abundant taxa in different fermented foods. A size-effect threshold of 2.0 on the logarithmic LDA score was used for discriminative functional biomarkers. On the species level, Bacillus subtilis, Tetragenococcus halophilus, and Clostridium arbusti were significantly more abundant in Doenjang than in Cheonggukjang, whereas Bacillus thermoamylovorans, Enterococcus faecium, and Lactobacillus sakei were significantly more abundant in Cheonggukjang than in Doenjang. Permutational multivariate analysis of variance (PERMANOVA) showed that the statistical difference in microbial clusters between the two groups was significant at the confidence level of p=0.001. This research could be used as basic research to identify the correlation between the biochemical characteristics of Korean fermented foods and the distribution of microbial communities.

• Korean Journal of Plant Resources
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• v.34 no.4
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• pp.278-286
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• 2021

Efficient in vitro regeneration system is essential for the successful crop breeding of soybean (Glycine max (L.) Merr.) using the new biotechnology. The genotype of donor plants strongly influences the establishment of tissue culture system. Therefore, the screening of genotypes with excellent tissue culture ability is very important for soybean genetic improvement. In this study, we report the tissue culture efficiency of 21 soybean cultivars belong to Korean soybean core-collection and two foreign cultivars (Jack and Maverick). The Kwangan, Anpyeong and Seonam are share close genetic relationship in 21 cultivars and these three cultivars were observed the high frequency of germination and regeneration. Furthermore, the high tissue culture abilities were also observed in the Williams 82 used in reference genome sequencing and the two foreign cultivars. The transformation of pBAtc:tRNA with bar gene was performed by Agrobacterium tumefaciens in the cultivars with high tissue culture ability. Transformation of the bar gene was identified by PCR analysis in Kwangan, Pungwon, Seonam, and Maverick. Our results provide useful information for the breeding of various soybean cultivars by plant biotechnology such as, genome editing.

• Korean Journal of Food Science and Technology
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• v.53 no.6
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• pp.775-784
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• 2021

After isolating the DIMO 52 strain with a large inhibition zone diameter for Clostridium perfringens and maximum butyric acid production from the fecal sample of a breastfeeding infant, it was identified as Clostidium butyricum. The maximum growth of the DIMO 52 strain was reached 24 h after inoculation, and the maximum butyric acid concentration was approximately 34.73±4.27 mM. The DIMO 52 strain survived approximately 67.5% of the initial inoculum at pH 2.0, and approximately 64.9% survived in RCM broth supplemented with 0.3% (w/v) oxgall. In addition, DIMO 52 showed antibacterial activity against Escherichia coli KCTC 2441 and Salmonella Typhimurium KCTC 1925. In LPS-stimulated RAW264.7 cells, 1×10³ CFU/mL viable cells of the DIMO 52 strain also exhibited significant NO (nitric oxide) production inhibitory activity (33%, p<0.01). This result suggests that C. butyricum DIMO 52 has anti-inflammatory activity related to NO radical-scavenging activity. In conclusion, C. butyricum DIMO 52 isolated in this study has the potential to be used as a probiotic.

• Journal of Microbiology and Biotechnology
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• v.29 no.7
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• pp.1083-1095
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• 2019

Butyrate is known to play a significant role in energy metabolism and regulating genomic activities that influence rumen nutrition utilization and function. Thus, this study investigated the effects of an isolated butyrate-producing bacteria, Clostridium saccharobutylicum, in rumen butyrate production, fermentation parameters and microbial population in Holstein-Friesian cow. An isolated butyrate-producing bacterium from the ruminal fluid of a Holstein-Friesian cow was identified and characterized as Clostridium saccharobutylicum RNAL841125 using 16S rRNA gene sequencing and phylogenetic analyses. The bacterium was evaluated on its effects as supplement on in vitro rumen fermentation and microbial population. Supplementation with $10^6CFU/ml$ Clostridium saccharobutylicum increased (p < 0.05) microbial crude protein, butyrate and total volatile fatty acids concentration but had no significant effect on $NH_3-N$ at 24 h incubation. Butyrate and total VFA concentrations were higher (p < 0.05) in supplementation with $10^6CFU/ml$ Clostridium saccharobutylicum compared with control, with no differences observed for total gas production, $NH_3-N$ and propionate concentration. However, as the inclusion rate (CFU/ml) of C. saccharobutylicum was increased, reduction of rumen fermentation values was observed. Furthermore, butyrate-producing bacteria and Fibrobacter succinogenes population in the rumen increased in response with supplementation of C. saccharobutylicum, while no differences in the population in total bacteria, protozoa and fungi were observed among treatments. Overall, our study suggests that supplementation with $10^6CFU/ml$ C. saccharobutylicum has the potential to improve ruminal fermentation through increased concentrations of butyrate and total volatile fatty acid, and enhanced population of butyrate-producing bacteria and cellulolytic bacteria F. succinogenes.

• Korean Journal of Microbiology
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• v.55 no.1
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• pp.74-76
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• 2019

Gram-positive anaerobic bacilli Actinomyces spp. commonly reside on mucosal surfaces of the oropharynx, gastrointestinal tract, and urogenital tract. Here, we first report the draft genome sequence of Actinomyces georgiae KHUD_A1, isolated from dental plaque of a Korean elderly woman. The genome is 2,652,059 bp in length and has a GC content of 68.06%. The genome includes 2,242 protein-coding genes, 9 rRNAs, and 64 tRNA. We identified 157 KHUD_A1 strain-specific genes, including genes encoding CPBP family intramembrane metalloprotease, bile acid: sodium symporter family protein, Txe/YoeB family addiction module toxin and Phd/YefM family antitoxin. The sequence information of A. georgiae KHUD_A1 will help understand the general characteristics of the bacterial species and the genome diversity of the genus Actinomyces.

• Journal of Microbiology and Biotechnology
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• v.31 no.9
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• pp.1231-1240
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• 2021

Members of the genus Bacillus are known to play an important role in promoting plant growth and protecting plants against phytopathogenic microorganisms. In this study, 21 isolates of Bacillus spp. were obtained from the root micro-ecosystem of Suaeda glauca. Analysis of the 16S rRNA genes indicated that the isolates belong to the species Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus subtilis, Bacillus pumilus, Bacillus aryabhattai and Brevibacterium frigoritolerans. One of the interesting findings of this study is that the four strains B1, B5, B16 and B21 are dominant in rhizosphere soil. Based on gyrA, gyrB, and rpoB gene analyses, B1, B5, and B21 were identified as B. amyloliquefaciens and B16 was identified as B. velezensis. Estimation of antifungal activity showed that the isolate B1 had a significant inhibitory effect on Fusarium verticillioides, B5 and B16 on Colletotrichum capsici (syd.) Butl, and B21 on Rhizoctonia cerealis van der Hoeven. The four strains grew well in medium with 1-10% NaCl, a pH value of 5-8, and promoted the growth of Arabidopsis thaliana. Our results indicate that these strains may be promising agents for the biocontrol and promotion of plant growth and further study of the relevant bacteria will provide a useful reference for the development of microbial resources.

• Korean Journal of Ecology and Environment
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• v.54 no.3
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• pp.247-256
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• 2021

Understanding the selective feeding behavior of zooplankton on phytoplankton is essential for evaluating the nutrient cycle and energy flow in the food web. Although many studies have been conducted regarding the feeding behaviors of zooplankton through gut content analyses, there are limitations in the visual identification of digested contents using a microscope. DNA techniques have been applied to overcome these limitations since they can detect and amplify small amounts of prey DNA remaining in the gut contents. We designed a method to extract prey DNA from the gut contents of the whole body of the copepod specimen and tested the resolution of DNA identification for the prey phytoplankton. The common brackish species, Sinocalanus tenellus, were collected from Saemangeum Reservoir in different sites and seasons, and gut content DNA was extracted using 2.5% bleach treatment for 2 min for removal of potential contamination sources existing in preserved specimens without dissolution of the body. The sequences of the extracted gut contents were confirmed using BLASTn suite based on the NCBI database. The phytoplankton species detected in the gut showed temporal and spatial differences. Although DNA analysis of small copepod gut contents has been suggested as an effective method to examine the dynamics of primary prey sources at the genus or species level, uncertainties such as misidentification and limitations in the detailed information of the composition still exist.

• Animal Bioscience
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• v.34 no.2
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• pp.243-255
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• 2021

Objective: Deoxynivalenol (DON) and zearalenone (ZEN) are mycotoxins that frequently contaminate maize and grain cereals, imposing risks to the health of both humans and animals and leading to economic losses. The gut microbiome has been shown to help combat the effects of such toxins, with certain microorganisms reported to contribute significantly to the detoxification process. Methods: We examined the cecum contents of three different dietary groups of pigs (control, as well as diets contaminated with 8 mg DON/kg feed or 0.8 mg ZEN/kg feed). Bacterial 16S rRNA gene amplicons were acquired from the cecum contents and evaluated by next-generation sequencing. Results: A total of 2,539,288 sequences were generated with ~500 nucleotide read lengths. Firmicutes, Bacteroidetes, and Proteobacteria were the dominant phyla, occupying more than 96% of all three groups. Lactobacillus, Bacteroides, Megasphaera, and Campylobacter showed potential as biomarkers for each group. Particularly, Lactobacillus and Bacteroides were more abundant in the DON and ZEN groups than in the control. Additionally, 52,414 operational taxonomic units were detected in the three groups; those of Bacteroides, Lactobacillus, Campylobacter, and Prevotella were most dominant and significantly varied between groups. Hence, contamination of feed by DON and ZEN affected the cecum microbiota, while Lactobacillus and Bacteroides were highly abundant and positively influenced the host physiology. Conclusion: Lactobacillus and Bacteroides play key roles in the process of detoxification and improving the immune response. We, therefore, believe that these results may be useful for determining whether disturbances in the intestinal microflora, such as the toxic effects of DON and ZEN, can be treated by modulating the intestinal bacterial flora.

• Animal Bioscience
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• v.34 no.1
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• pp.93-101
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• 2021

Objective: Temperature could influence protein and amino acid deposition as well as gut microbiota profile and composition. However, the specific effects of ambient temperature on amino acids deposition and gut microbiota composition remain insufficiently understood. Methods: A total of 300 one-day-old Avian broilers were randomly divided into three groups and reared at high, medium, and low temperature (HT, MT, and LT), respectively. Breast muscle and fecal samples were collected for amino acid composition analysis and 16S rRNA gene sequence analysis. Results: Our data showed that compared to the MT group, there was a decrease of muscle leucine and tyrosine (p<0.05), as well as an increase of methionine in the HT group (p<0.05) and a decrease of serine in the LT group. Examination of microbiota shift revealed that at genus level, the relative abundance of Turicibacter and Parabacteroides was increased in the HT group (p<0.05) and that the relative abundances of Pandoraea, Achromobacter, Prevotella, Brevundimonas, and Stenotrophomonas in the LT group were higher than those in the MT group (p<0.05). In addition, there were substantial correlations between microbes and amino acids. In the HT group. Turicibacter was negatively correlated with aspartic acid and tyrosine, whereas Parabacteroides was positively correlated with methionine (p<0.05). In the LT group, there were multiple positive correlations between Achromobacter and arginine, isoleucine or tyrosine; between Prevotella and cysteine or phenylalanine; between Brevundimonas and cysteine; and between Stenotrophomonas and cysteine as well as a negative correlation between Stenotrophomonas and serine. Conclusion: Our findings demonstrated that amino acid content of breast muscle and intestinal microbiota profile was affected by different ambient temperatures. Under heat exposure, augmented abundance of Parabacteroides was correlated with elevated methionine. Low temperature treatment may affect muscle tyrosine content through the regulation of Achromobacter.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.4
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• pp.297-306
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• 2020

The aims of the present study were to confirm that regulation of the PA and environment via TGF-β regulation of sperm by Percoll-separated in porcine uterine epithelial cells. And, it was performed to identify the cytokines (TGF-β1, 2 and 3, TGF-β receptor1 and 2; interleukin, IL-6, IL-8) and PA-related genes (urokinase-PA, uPA; tissue-PA, tPA; PA inhibitor, PAI; uPA-receptor, uPAR) by spermatozoa. The experiment used porcine uterus epithelial cells (pUECs) and uterine tissue epithelial cells, Boar sperm were separated by discontinuous Percoll density gradient (45/90%), and tissues were co-incubated with spermatozoa, followed by real-time PCR. PA activity was measured of sperm by discontinuous Percoll density gradient (45/90%) for 24 hours. To measure viability and acrosome damage of sperm double stained propidium iodide (PI) and SYBR-14 or FITC-PNA were used. In results, binding ratio of Percoll-separated sperm was found no differences, but sperms isolated from 90% Percoll layer reduced PA activity (p < 0.05). when co-cultured sperm selected Percoll in porcine uterus tissues epithelial cells, 90% layer sperm increased TGF-β R1, contrastively tPA and PAI-1 in comparison with control (p < 0.05). 45% sperm was decreased the expression of uPA (p < 0.05). TGF-β decreased PA activity in the supernatant collected from pUECs (p < 0.05). Especially, The group including uPA, PAI-1 were induce sperm intact, while it was reduced in sperm damage when compared to control (p < 0.05). Also, there was no significant difference group of tPA and tPA+I in the dead sperm and acrosome damage compared to control. The expression of tPA and PAI showed a common response. Percoll-separated spermatozoa in 90% layer reduced tPA and IL-related gene mRNA expression. Thus, Percoll-sparated sperm in 90% layer show that it can suppress inflammation through increased expression of TGF-β and downregulation of PA and IL in epithelial cells compared to 45% layer Percoll.