• Journal of Embryo Transfer
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• v.21 no.4
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• pp.345-351
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• 2006

The objective of this study was to investigate the effects of preservation period of porcine liquid semen on bacterial contamination and in vitro production of embryo. Extended liquid semen was prepared by three mixture of boar's ejaculates from each farm without antibiotics, and were kept in $17^{\circ}C$ semen preservation incubator until use. Sperm motility was significantly (p<0.05) decreased as semen preservation time goes by (78.7$\pm$2.4% for 1 day vs. 71.1$\pm$2.4 and 64.8$\pm$2.4% for 3 and 5 days of presentation, respectively). Quantitative of bacteria in semen was significantly (p<0.05) higher in 5 days ($57.8\pm105.2\times10^4$ Cfu) compared to 0 and 3 days ($32.1\pm76.8\times10^4$ and $26.9\pm46.6\times10^4$ Cfu, respectively) of preservation. In terms of development of in vitro fertilization of porcine embryos inseminated by preserved semen, the rate of normal fertilization (2PN) was significantly (p<0.05) decreased in 5 days (56.0$\pm$2.6%) compared to 1 and 3 days (66.0$\pm$2.7 and 64.0$\pm$2.7%, respectively) of preservation. Cleavage rate was also significantly (p<0.05) affected by preservation period (75.0$\pm$4% for 1 day, 70.0$\pm$0.3 and 71.0$\pm$0.3% for 3 and 5 days, respectively). The in vitro developmental rate of blastocyst stage embryo was significantly (p<0.05) affected by semen preservation period (15.0$\pm$1.0% for 1 day vs. 11.0$\pm$0.9 and 8.0$\pm$0.9% for 3 and 5 days, respectively). It is concluded that more than 3 days of liquid semen preservation without antibiotics increased the quantity of bacteria resulted in detrimental effect on sperm motility and decreased both normal insemination rate and the developmental rate of blastocyst stage embryo.

• Journal of Life Science
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• v.22 no.2
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• pp.259-265
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• 2012

A bacterial strain, CK214, exhibiting high motility on an LB agar (1.5%, w/v) surface was isolated from the environment. The formation of unusual agar shrinking around colonies on agar plates was observed. The strain grew on minimal media containing pure agar as a sole carbon source. The cell-free culture supernatant of CK214 generated a reduced form of sugar in the in vitro reaction with the use of pure agar as a substrate, suggesting the secretion of an agar-degrading enzyme. The CK214 strain showed swarming motility on the solid media containing a wide range of concentrations of agar (0.5, 1.0, 1.5, 2.0% w/v). Various tests, including Gram staining, API analysis, and phylogenetic analysis based on 16S rDNA sequences identified that the CK214 strain was a G(+) rod-shaped bacterium grouped in genus Paenibacillus. Electron microscopic analysis demonstrated that the P. CK214 strain is peritrichously flagellated. Through transposon random mutagenesis, several agar-degrading activity defective mutants (ADMs) were generated. These mutants will be used in the future experimentation for the study of the correlation between agar-degrading activity and motility.

• Journal of Microbiology and Biotechnology
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• v.24 no.12
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• pp.1609-1621
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• 2014

The plant pathogen Burkholderia gladioli, which has a broad host range that includes rice and onion, causes bacterial panicle blight and sheath rot. Based on the complete genome sequence of B. gladioli BSR3 isolated from infected rice sheaths, the genome of B. gladioli BSR3 contains the luxI/luxR family of genes. Members of this family encode N-acyl-homoserine lactone (AHL) quorum sensing (QS) signal synthase and the LuxR-family AHL signal receptor, which are similar to B. glumae BGR1. In B. glumae, QS has been shown to play pivotal roles in many bacterial behaviors. In this study, we compared the QS-dependent gene expression between B. gladioli BSR3 and a QS-defective B. gladioli BSR3 mutant in two different culture states (10 and 24 h after incubation, corresponding to an exponential phase and a stationary phase) using RNA sequencing (RNA-seq). RNA-seq analyses including gene ontology and pathway enrichment revealed that the B. gladioli BSR3 QS system regulates genes related to motility, toxin production, and oxalogenesis, which were previously reported in B. glumae. Moreover, the uncharacterized polyketide biosynthesis is activated by QS, which was not detected in B. glumae. Thus, we observed not only common QS-dependent genes between B. glumae BGR1 and B. gladioli BSR3, but also unique QS-dependent genes in B. gladioli BSR3.

• KSBB Journal
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• v.29 no.3
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• pp.131-138
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• 2014

In this study, we isolated two novel chitinase producing bacterial strains from yellow loess samples collected from Jullanamdo province. The chitinase producing bacteria were isolated based on the zone size of clearance in the chitin agar plates. Both of them were gram positive, rod ($2{\sim}3{\times}0.3{\sim}0.4{\mu}m$), spore-forming, and motility positive. They were facultative anaerobic, catalase positive and hydrolyzed starch, gelatin, and casein. From the 16s rRNA gene sequence analysis, the isolates were labeled as Bacillus licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02. The isolates showed higher extracellular chitinase activities than B. licheniformis ATCC 14580 as a control. The optimum temperature and pH for chitinase production were $40^{\circ}C$ and pH 7.0, respectively. Response Surface Methodology (RSM) was used to optimize the culture medium for efficient production of the chitinase. Under this optimal condition, 1.5 times higher chitinase activity of B. licheniformis KYLS-CU02 was obtained. Extracellular chitinases of the two isolates were purified through ammonium sulfate precipitation and anion-exchange DEAE-cellulose column chromatography. The specific activities of purified chitinase from B. licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02 were 7.65 and 5.21 U/mg protein, respectively. The molecular weights of the two purified chitinases were 59 kDa. Further, the purified chitinase of B. licheniformis KYLS-CU01 showed high antifungal activity against Fusarium sp.. In conclusion, these two bacterial isolates can be used as a biopesticide to control pathogenic fungi.

• KSBB Journal
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• v.31 no.3
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• pp.158-164
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• 2016

Plant growth promoting (PGP) hormones, which are produced in a small quantity by bacteria, affect in plant growth and development. PGPs play an important role on the crop productivity in agricultural field. In this study, a photosynthetic bacterial strain producing the PGP was isolated from paddy soil. Bacterial isolate was gram negative, rod-shaped and motility positive. From the 16s rRNA gene sequence analysis, the isolate was identified as Rhodobacter capsulatus PS-2. The mass cultivation of R. capsulatus PS-2 was optimized by considering of the carbon, nitrogen and inorganic salt sources. Optimal medium composition was determined as Na-succinate 4.5 g, yeast extract 5 g, $K_2HPO_4$ 1 g, $MgSO_4$ 5 g, per liter. From the result of 500 L fermentation for 2 days using the optimal medium, the viable cells were $8.7{\times}10^9cfu/mL$. R. capsulatus PS-2 strain produced the carotenoid and indole-3-acetic acid (IAA). The carotenoid extraction and quantitative analysis were performed by HCl-assisting method. Total carotenoid contents from R. capsulatus PS-2 culture broth were measured as $7.02{\pm}0.04$ and $6.93{\pm}0.05mg/L$ under photoheterotrophic and chemoheterotrophic conditions, respectively. To measure the productivity of IAA, colorimetric method was employed using Salkowski reagent at optical density 535 nm. The results showed that the highest content of IAA was $197.44{\pm}5.92mg/L$ in the optimal medium supplemented with 0.3% tryptophan.

• Transactions of the Korean Society of Mechanical Engineers B
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• v.31 no.3 s.258
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• pp.209-216
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• 2007

Bacterial chemotaxis is essential to the study of structure and function of bacteria. Although many studies have accumulated the knowledge about chemotaxis in the past, the motion of a single bacterium has not been studied much yet. In this study, we have developed a device microfabricated by soft lithography and consisting of microfluidic channels. The microfluidic assay generates a concentration gradient of chemoattractant linearly in the main channel by only diffusion of the chemicals. Bacteria are injected into the main channel in a single row by hydrodynamic focusing technique. We measured the velocity of bacteria in response to a given concentration gradient of chemoattractant using the microfludic assay, optical systems with CCD camera and simple PTV (Particle Tracking Velocimetry) algorithm. The advantage of this assay and experiment is to measure the velocity of a single bacterium and to quantify the degree of chemotaxis by statistically analyzing the velocity at the same time. Specifically, we measured and analyzed the motility of Escherichia coli strain RP437 in response to various concentration gradients of L-aspartate statistically and quantitatively by using this microfluidic assay. We obtained the probability density of the velocity while RP437 cells are swimming and tumbling in the presence of the linear concentration gradient of L-aspartate, and quantified the degree of chemotaxis by analyzing the probability density.

• Journal of Microbiology and Biotechnology
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• v.30 no.11
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• pp.1729-1738
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• 2020

Salmonellosis is a form of gastroenteritis caused by Salmonella infection. The main transmission route of salmonellosis has been identified as poorly cooked meat and poultry products contaminated with Salmonella. However, in recent years, the number of outbreaks attributed to contaminated raw produce has increased dramatically. To understand how Salmonella adapts to produce, transcriptomic analysis was conducted on Salmonella enterica serovar Virchow exposed to fresh-cut radish greens. Considering the different Salmonella lifestyles in contact with fresh produce, such as motile and sessile lifestyles, total RNA was extracted from planktonic and epiphytic cells separately. Transcriptomic analysis of S. Virchow cells revealed different transcription profiles between lifestyles. During bacterial adaptation to fresh-cut radish greens, planktonic cells were likely to shift toward anaerobic metabolism, exploiting nitrate as an electron acceptor of anaerobic respiration, and utilizing cobalamin as a cofactor for coupled metabolic pathways. Meanwhile, Salmonella cells adhering to plant surfaces showed coordinated upregulation in genes associated with translation and ribosomal biogenesis, indicating dramatic cellular reprogramming in response to environmental changes. In accordance with the extensive translational response, epiphytic cells showed an increase in the transcription of genes that are important for bacterial motility, nucleotide transporter/metabolism, cell envelope biogenesis, and defense mechanisms. Intriguingly, Salmonella pathogenicity island (SPI)-1 and SPI-2 displayed up- and downregulation, respectively, regardless of lifestyles in contact with the radish greens, suggesting altered Salmonella virulence during adaptation to plant environments. This study provides molecular insights into Salmonella adaptation to plants as an alternative environmental reservoir.

• Journal of Ginseng Research
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• v.45 no.1
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• pp.75-85
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• 2021

Background: Invasive infections due to foodborne pathogens, including Salmonella enterica serovar Typhimurium, are prevalent and life-threatening. This study aimed to evaluate the effects of ginsenoside Rg3 (Rg3) on the adhesion, invasion, and intracellular survival of S. Typhimurium. Methods: The impacts of Rg3 on bacterial growth and host cell viability were determined using the time kill and the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assays, respectively. Gentamicin assay and confocal microscopic examination were undertaken to determine the effects of Rg3 on the adhesive and invasive abilities of S. Typhimurium to Caco-2 and RAW 264.7 cells. Quantitative reverse transcription polymerase chain reaction was performed to assess the expression of genes correlated with the adhesion, invasion, and virulence of S. Typhimurium. Results: Subinhibitory concentrations of Rg3 significantly reduced (p < 0.05) the adhesion, invasion, and intracellular survival of S. Typhimurium. Rg3 considerably reduced (p < 0.05) the bacterial motility as well as the release of nitrite from infected macrophages in a concentration-dependent manner. The expression of genes related to the adhesion, invasion, quorum sensing, and virulence of S. Typhimurium including cheY, hilA, OmpD, PrgK, rsgE, SdiA, and SipB was significantly reduced after Rg3 treatment. Besides, the compound downregulated rac-1 and Cdc-42 that are essential for actin remodeling and membrane ruffling, thereby facilitating Salmonella entry into host cells. This report is the first to describe the effects of Rg3 on "trigger" entry mechanism and intracellular survival S. Typhimurium. Conclusion: Rg3 could be considered as a supplement agent to prevent S. Typhimurium infection.

• 한국전산유체공학회:학술대회논문집
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• 2009.11a
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• pp.164-169
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• 2009

Study on swimming of microorganisms like, sperm motility, cilia beating, bacterial flagellar propulsion has found immense significance in the field of biological fluiddynamics. Because of the complexity involved, it is challenging for the researchers to model such problems. Immersed boundary method has proved its efficacy in the field of biological fluiddynamics, The present work aims at performing a numerical study on the microorganism locomotion using the immersed boundary method proposed by Peskin[1]. A two-dimensional model of the microorganism is modeled as thin elastic filament described as a sine wave. The neutrally buoyant organism undergoing deformations is immersed in a viscous and incompressible fluid. The fluid quantities are described using Eulerian coordinates and the immersed body is represented by Lagrangian coordinates. The Eulerian and Lagrangian variables are connected by the Dirac delta function. The Navier-Stokes equations governing the fluid flow are solved using the fractional step method on a staggered Cartesian grid system. The developed numerical code in FORTRAN will be validated by comparing the numerical results with the available results.

• Microbiology and Biotechnology Letters
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• v.47 no.4
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• pp.651-661
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• 2019

Targeting the pathogen viability using drugs is associated with development of drug resistance due to selective pressure. Hence, there is an increased interest in developing agents that target bacterial virulence. In this study, the inhibitory effect of ciclopirox, an antifungal agent with iron chelation potential, on the microbial virulence factors was evaluated in 26 clinical MDR Pseudomonas aeruginosa isolates collected from Alexandria Main University Hospital, a tertiary hospital in Egypt. Treatment with 9 ㎍/ml ciclopirox inhibited the hemolytic activity in 70% isolates, reduced pyocyanin production, decreased protease secretion in 46% isolates, lowered twitching and swarming motility, and decreased biofilm formation by 1.5- to 4.5-fold. The quantitative real-time PCR analysis revealed that treatment with ciclopirox downregulated the expression levels of alkaline protease (aprA) and pyocyanin (phzA1). Ciclopirox is used to treat hematological malignancies and the systemic administration of ciclopirox is reported to have adequate oral absorption with a satisfactory drug safety profile. It is important to calculate the appropriate clinical dose and therapeutic index to reposition ciclopirox from a topical antifungal agent to a promising virulence-modifying agent agent against P. aeruginosa, a problematic Gram-negative pathogen.