• Journal of Microbiology and Biotechnology
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• v.33 no.1
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• pp.83-95
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• 2023

These days, bacterial detection methods have some limitations in sensitivity, specificity, and multiple detection. To overcome these, novel detection and identification method is necessary to be developed. Recently, NGS panel method has been suggested to screen, detect, and even identify specific foodborne pathogens in one reaction. In this study, new NGS panel primer sets were developed to target 13 specific virulence factor genes from five types of pathogenic Escherichia coli, Listeria monocytogenes, and Salmonella enterica serovar Typhimurium, respectively. Evaluation of the primer sets using singleplex PCR, crosscheck PCR and multiplex PCR revealed high specificity and selectivity without interference of primers or genomic DNAs. Subsequent NGS panel analysis with six artificially contaminated food samples using those primer sets showed that all target genes were multi-detected in one reaction at 10⁸-10⁵ CFU of target strains. However, a few false-positive results were shown at 10⁶-10⁵ CFU. To validate this NGS panel analysis, three sets of qPCR analyses were independently performed with the same contaminated food samples, showing the similar specificity and selectivity for detection and identification. While this NGS panel still has some issues for detection and identification of specific foodborne pathogens, it has much more advantages, especially multiple detection and identification in one reaction, and it could be improved by further optimized NGS panel primer sets and even by application of a new real-time NGS sequencing technology. Therefore, this study suggests the efficiency and usability of NGS panel for rapid determination of origin strain in various foodborne outbreaks in one reaction.

• Animal Bioscience
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• v.36 no.8
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• pp.1241-1251
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• 2023

Objective: Egg yolk immunoglobulin (IgY) is an antibiotic alternative to prevent and fight intestinal pathogenic infections. This study aimed to investigate the effects of sodium alginate/chitosan/sodium alginate IgY microcapsules on the growth performance, serum parameters, and intestinal health of broiler chickens. Methods: One-day-old broilers (Ross 308) were divided into five treatments, each with 10 replicates of five chickens. The dietary treatments were maintained for 28 days and consisted of a basal diet (NC), basal diet + 500 mg chlortetracycline/kg diet (CH), basal diet + 50 mg non-microencapsulated IgY/kg diet (NM), basal diet + 600 mg low levels microencapsulated IgY/kg diet (LM), and basal diet + 700 mg high levels microencapsulated IgY/kg diet (HM). Results: Throughout the 28-day trial period, the NM, LM, HM, and CH groups increased average daily gain compared with the NC group (p<0.05), and the HM group reduced feed conversion ratio compared with the CH group (p<0.05). The LM and HM groups increased relative organ weights of thymus and spleen compared with the CH and NM groups (p<0.05). The HM group improved the duodenal, jejunal and ileum villi height (VH) and villus height to crypt depth ratio (VH:CD) compared with the CH and NM groups (p<0.05). Compared with the CH group, the HM group increased serum immunoglobulin (IgA), immunoglobulin G (IgG), superoxide dismutase, total antioxidant capacity, and glutathione peroxidase levels (p<0.05), and decreased serum malondialdehyde levels (p<0.05). Compared with the NC group, the NM, LM, HM, and CH groups reduced colonic Escherichia coli and Salmonella levels (p<0.05). and the HM group promoted the levels of lactic acid bacteria and bifidobacteria compared with the CH group (p<0.05). Conclusion: Microencapsulation could be considered as a way to improve the efficiency of IgY. The 700 mg high levels microencapsulated IgY/kg diet could potentially be used as an alternative to antibiotics to improve the immune performance and intestinal health, leading to better performance of broiler chickens.

• Korean Journal of Environmental Biology
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• v.39 no.4
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• pp.445-451
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• 2021

In this study, we described the production of an antibody to living modified organisms (LMOs) containing the gene encoding for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium tumefaciens strain CP4 EPSPS provides resistance to the herbicide glyphosate (N- (phosphonomethyl) glycine). These LMOs were approved and have recently been used in the feed, food production, and processing industries in South Korea. Highly efficient monoclonal antibody (mAb) production is crucial for developing assays that enable the proper detection and quantification of the CP4 EPSPS protein in LMOs. This study describes the purification and characterization of recombinant CP4 EPSPS protein in E. coli BL21 (DE3) based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrixassisted laser desorption/ionization time-of-flight mass spectrometry. The production of mAbs was undertaken based on the standard operating procedure of Abclon, Inc.(South Korea), and the purity of the mAbs was assessed using SDS-PAGE. The following five mAb clones were produced: 2F2, 4B9, 6C11, 10A9, and 10G9. To verify the efficiency and specificity of the five developed mAbs, we performed Western blotting analysis using the LM (living modified) cotton crude extracts. All mAbs could detect the CP4 EPSPS protein in the LM cotton traits MON1445 and MON88913 with high specificity, but not in any other LM cottons or non-LM cottons. These data indicate that these five mAbs to CP4 EPSPS could be successfully used for the further development of antibody-based detection methods to target CP4 EPSPS protein in LMOs.

• Korean Journal of Food Science and Technology
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• v.54 no.1
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• pp.80-87
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• 2022

We investigated the effects of organic acid and ultrasonication treatment in maintaining the quality of garlic during storage. Samples were exposed for 5 min to either ultrasonication at 60℃, 1% citric acid, or 0.5% fumaric acid. Presence of microorganisms and minerals, hardness, and color were compared during storage at 4℃ for 28 days. The total aerobic bacterial count remained low. No proliferation of Escherichia coli was observed after treatment with fumaric acid or ultrasonication, and mold proliferation was inhibited by ultrasonication. The mineral content of the northern type garlic was higher than that of the southern type. Exposure to fumaric acid did not result in a substantial difference in hardness until 21 days of storage, at which time there was a decrease in the L-value in each cultivation type. Our results indicate that treatment with 0.5% fumaric acid for 5 min was effective in reducing the abundance of microorganisms during storage without affecting the hardness or color in garlic.

• Korean journal of applied entomology
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• v.62 no.2
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• pp.95-102
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• 2023

The number of households with pets has increased in recent years. Consequently, interest in mites and their odors has increased. In this study, we investigated whether pyrethrum extract and water-soluble phytoncides are effective tick repellents, deodorizing agents, and antibacterial agents. In the tick-climbing test, the pyrethrum extract (23.33 points) and water-soluble phytoncide (22.00 points) showed a high repelling effect. To confirm the tick avoidance effect of the composite extract, a tick climbing test was conducted with a pest control extract, a composite extract (referred to as a sample) containing a water-soluble phytoncide, and a third-party product (control). The sample showed a high avoidance effect (13.00 points and Control 26.67 points. An indoor repellent efficacy test for ticks confirmed that the sample showed an excellent tick-repellent effect (88%) compared to the control (12%). As a result of a deodorization test, ammonia was shown to have a deodorizing effect of 98.3% and trimethylamine of 99.5%, whereas, in the antibacterial test, the E. coli, Staphylococcus aureus, and pneumonia groups showed an antibacterial effect of 99.9%. Combining the results of this study, it is judged that the complex extract including the above material has high utility value as a material that is effective in mite repellents, deodorization, and antibacterial activity.

• Journal of Wetlands Research
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• v.25 no.4
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• pp.326-334
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• 2023

The purpose of this study is to use long-term water quality monitoring data from tributaries of the Nakdong River system to identify problematic substances in tributaries by examining the rate of exceedance and increase in water quality targets. In the Nakdong River system, monitoring is conducted once a month for 38 tributaries that require intensive management, and this data was used to analyze trends in exceeding and increasing target water quality at each point. The analysis items are eight items that can be evaluated based on river water quality standards: DO, BOD, COD, TOC, SS, total phosphorus, fecal coliform, and total coliform. As a result of the analysis, the target water quality exceedance rate was more than 50%, and the items with an increasing trend were TOC, fecal coliform and total E. coli counts, and the items with an exceedance rate of less than 50% but an increasing trend were SS. TOC is believed to be caused by an increase in non-degradable substances, and the continued increase in Total Coliform will require management of Total ColiformTotal Coliform in effluent water from sewage treatment facilities in the future.

• Journal of the Korean Applied Science and Technology
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• v.41 no.1
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• pp.1-8
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• 2024

The safety of cosmetic ingredients is considered paramount. In order to enhance safety, a novel preservative, PE-gal, was bio-synthesized by utilizing the Escherichia coli enzyme 𝛽-galactosidase on the conventional preservative 2-phenoxyethanold (PE). The skin absorption of the bio-synthesized product, PE-gal, intended for use in cosmetics, was evaluated for permeability using the Franz Diffusion Cell Assay system, comparing it with the conventinal preservative PE. When using samples of the same mass concentration, the Flux and Kp values of PE increased over time, indicating a gradual increase in permeability. However, PE-gal did not exhibit sufficient permeability to measure. This suggests that the skin permeability of PE is higher than that of the PE-gal saccharide. According to Marzulli et al., when confirming the degree of permeation using Kp values, the permeation rate of PE was measured as "slow" at a concentration of 1mg/mL. Thus, the transdermal permeability of the divedened form of PE-gal was significantly lower compared to PE.

• Journal of Applied Biological Chemistry
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• v.66
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• pp.519-526
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• 2023

Present study explored immunostimulatory effects of Astragalus membranaceus and Zanthoxylum schinifolium 1:1 (w:w) mixture (AZM-1:1) in Raw264.7 cells, mouse macrophage derived cells. Treatment with 100-400 ㎍/mL of AZM-1:1 in Raw264.7 cells significantly increased nitric oxide production in parallel with inducible nitric oxide synthase mRNA expression without affecting cytotoxicity. In addition, AZM-1:1 dose-dependently increased prostaglandin E₂ production in conditioned medium along with cyclooxygenase-2 mRNA induction. Moreover, AZM-1:1 induces the transcription of tumor necrosis factor-α, interleukin-1β, interleukin-6, and monocyte chemoattractant protein-1. Immunoblot analyses revealed that AZM-1:1 significantly increased the phosphorylation of mitogen-activated protein kinases, provoked phosphorylation-mediated degradation of inhibitory-κBα, and phosphorylated p65. Furthermore, treatment with AZM-1:1 promoted phagocytosis of Escherichia coli particle labeled with green fluorescence. Taken together, AZM-1:1 may be a promising nutraceutical for stimulation the innate immune system, including macrophages.

• Animal Bioscience
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• v.37 no.4
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• pp.689-696
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• 2024

Objective: Our study examined the impact of propriety blends of Bacillus strain probiotics on the performance, egg quality, and faecal microflora of laying hens. Methods: A total of 183 Institut de selection Animale (ISA) brown laying hens aged 23 weeks with an average body weight of 1,894±72 g were randomly allocated into 3 groups as control (corn-soybean meal based diet, CON), 0.5 g/kg Enterosure probiotics (ET1, 3×10⁸ colony-forming unit [CFU]/kg feed), and 5 g/kg Enterosure probiotics (ET2, 3×10⁹ CFU/kg feed) administered in mashed form. At the completion of each phase hen day egg production (HDEP), average egg weight (AEW), feed intake, and faecal microbiota were evaluated. Results: HDEP and AEW were higher (p<0.05) in the ET2-supplemented diet in phase 3 (week 9 to 12) compared with CON. Egg mass (EM) was higher (p<0.05) in phase 2 at ET2, and also higher (p<0.05) in phase 3 at the ET1 and ET2-supplemented diets compared with CON. Feed conversion ratio was lower (p<0.05) in phase 3 at the ET1 and ET2-supplemented diets, with ET2 being the lowest compared with ET1 and CON. Yolk colour was higher (p<0.05) in the ET-supplemented diets at phase 3 compared with CON. Bifidobacterium spp. was higher (p<0.05) in the ET2- supplemented diet compared with CON in phase 2, while in In phase 3, Lactobacillus spp. and Bifidobacterium spp. were higher (p<0.05) in the ET-supplemented diets compared with CON. Coliforms were lower (p<0.05) in the ET-supplemented diets compared with CON in phase 3. Conclusion: The propriety blends of Bacillus strain probiotics supplements at 0.5 g/kg and 5 g/kg could improve the production and quality of eggs with more significance at 5 g/kg for HDEP, AEW and EM, which was achieved via the increase of beneficial microbiomes such as Lactobacillus spp., Bifidobacterium spp., and the decrease of pathogenic microbiomes like Escherichia coli and Coliforms which was speculated to improve gut barrier function and the reproductive hormone.

• IMMUNE NETWORK
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• v.16 no.5
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• pp.296-304
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• 2016

It has been reported that fatty acid binding proteins (FABPs) do not act only as intracellular mediators of lipid responses but also have extracellular functions. This study aimed to investigate whether extracellular liver type (L)-FABP has a biological activity and to determined serum L-FABP levels in patients with end-stage renal disease (ESRD). We isolated L-FABP complementary deoxyribonucleic acid (cDNA) from the Huh7 human hepatocarcinoma cell line and expressed the recombinant L-FABP protein in Escherichia coli. A549 lung carcinoma and THP-1 monocytic cells were stimulated with the human recombinant L-FABP. Human whole blood cells were also treated with the human recombinant L-FABP or interleukin (IL)-1α. IL-6 levels were measured in cell culture supernatants using IL-6 enzyme-linked immunosorbent assay (ELISA). Human recombinant L-FABP induced IL-6 in a dose-dependent manner in A549, THP-1 cells, and whole blood cells. The blood samples of healthy volunteers and patients with ESRD were taken after an overnight fast. The serum levels of L-FABP in healthy volunteers and ESRD patients were quantified with L-FABP ELISA. The values of L-FABP in patients with ESRD were significantly lower than those in the control group. Our results demonstrated the biological activity of L-FABP in human cells suggesting L-FABP can be a mediator of inflammation.