• Title/Summary/Keyword: Strain Concentration

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Identification of N,N',N"-triacetylfusarinine C as a key metabolite for root rot disease virulence in American ginseng

  • Walsh, Jacob P.;DesRochers, Natasha;Renaud, Justin B.;Seifert, Keith A.;Yeung, Ken K.C.;Sumarah, Mark W.
    • Journal of Ginseng Research
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    • v.45 no.1
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    • pp.156-162
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    • 2021
  • Background: It is estimated that 20-30% of ginseng crops in Canada are lost to root rot each harvest. This disease is commonly caused by fungal infection with Ilyonectria, previously known as Cylindrocarpon. Previous reports have linked the virulence of fungal disease to the production of siderophores, a class of small-molecule iron chelators. However, these siderophores have not been identified in Ilyonectria. Methods: High-resolution LC-MS/MS was used to screen Ilyonectria and Cylindrocarpon strain extracts for secondary metabolite production. These strains were also tested for their ability to cause root rot in American ginseng and categorized as virulent or avirulent. The differences in detected metabolites between the virulent and avirulent strains were compared with a focus on siderophores. Results: For the first time, a siderophore N,N',N"-triacetylfusarinine C (TAFC) has been identified in Ilyonectria, and it appears to be linked to disease virulence. Siderophore production was suppressed as the concentration of iron increased, which is in agreement with previous reports. Conclusion: The identification of the siderophore produced by Ilyonectria gives us further insight into the root rot disease that heavily affects ginseng crop yields. This research identifies a molecular pathway previously unknown for ginseng root rot and could lead to new disease treatment options.

4,4'-Diaponeurosporene from Lactobacillus plantarum subsp. plantarum KCCP11226: Low Temperature Stress-Induced Production Enhancement and In Vitro Antioxidant Activity

  • Kim, Mibang;Jung, Dong-Hyun;Seo, Dong-Ho;Park, Young-Seo;Seo, Myung-Ji
    • Journal of Microbiology and Biotechnology
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    • v.31 no.1
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    • pp.63-69
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    • 2021
  • Carotenoids, which have biologically beneficial effects and occur naturally in microorganisms and plants, are pigments widely applied in the food, cosmetics and pharmaceutical industries. The compound 4,4'-diaponeurosporene is a C30 carotenoid produced by some Lactobacillus species, and Lactobacillus plantarum is the main species producing it. In this study, the antioxidant activity of 4,4'-diaponeurosporene extracted from L. plantarum subsp. plantarum KCCP11226 was examined. Maximum carotenoid content (0.74 ± 0.2 at A470) was obtained at a relatively low temperature (20℃). The DPPH radical scavenging ability of 4,4'-diaponeurosporene (1 mM) was approximately 1.7-fold higher than that of butylated hydroxytoluene (BHT), a well-known antioxidant food additive. In addition, the ABTS radical scavenging ability was shown to be 2.3- to 7.5-fold higher than that of BHT at the range of concentration from 0.25 mM to 1 mM. The FRAP analysis confirmed that 4,4'-diaponeurosporene (0.25 mM) was able to reduce Fe3+ by 8.0-fold higher than that of BHT. Meanwhile, 4,4'-diaponeurosporene has been confirmed to be highly resistant to various external stresses (acid/bile, high temperature, and lysozyme conditions). In conclusion, L. plantarum subsp. plantarum KCCP11226, which produces 4,4'-diaponeurosporene as a functional antioxidant, may be a potentially useful strain for the development of functional probiotic industries.

Antimicrobial Resistance Profile of Acinetobacter spp. Isolates from Retail Meat Samples under Campylobacter-Selective Conditions

  • Cha, Min-Hyeok;Kim, Sun Hee;Kim, Seokhwan;Lee, Woojung;Kwak, Hyo-Sun;Chi, Young-Min;Woo, Gun-Jo
    • Journal of Microbiology and Biotechnology
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    • v.31 no.5
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    • pp.733-739
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    • 2021
  • Acinetobacter strains are widely present in the environment. Some antimicrobial-resistant strains of this genus have been implicated in infections acquired in hospitals. Genetic similarities have been reported between Acinetobacter strains in nosocomial infections and those isolated from foods. However, the antimicrobial resistance of Acinetobacter strains in foods, such as meat, remains unclear. This study initially aimed to isolate Campylobacter strains; instead, strains of the genus Acinetobacter were isolated from meat products, and their antimicrobial resistance was investigated. In total, 58 Acinetobacter strains were isolated from 381 meat samples. Of these, 32 strains (38.6%) were from beef, 22 (26.5%) from pork, and 4 (4.8%) from duck meat. Antimicrobial susceptibility tests revealed that 12 strains were resistant to more than one antimicrobial agent, whereas two strains were multidrug-resistant; both strains were resistant to colistin. Cephalosporin antimicrobials showed high minimal inhibitory concentration against Acinetobacter strains. Resfinder analysis showed that one colistin-resistant strain carried mcr-4.3; this plasmid type was not confirmed, even when analyzed with PlasmidFinder. Analysis of the contig harboring mcr-4.3 using BLAST confirmed that this contig was related to mcr-4.3 of Acinetobacter baumannii. The increase in antimicrobial resistance in food production environments increases the resistance rate of Acinetobacter strains present in meat, inhibits the isolation of Campylobacter strains, and acts as a medium for the transmission of antimicrobial resistance in the environment. Therefore, further investigations are warranted to prevent the spread of antimicrobial resistance in food products.

Simulation study on the mechanical properties and failure characteristics of rocks with double holes and fractures

  • Pan, Haiyang;Jiang, Ning;Gao, Zhiyou;Liang, Xiao;Yin, Dawei
    • Geomechanics and Engineering
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    • v.30 no.1
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    • pp.93-105
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    • 2022
  • With the exploitation of natural resources in China, underground resource extraction and underground space development, as well as other engineering activities are increasing, resulting in the creation of many defective rocks. In this paper, uniaxial compression tests were performed on rocks with double holes and fractures at different angles using particle flow code (PFC2D) numerical simulations and laboratory experiments. The failure behavior and mechanical properties of rock samples with holes and fractures at different angles were analyzed. The failure modes of rock with defects at different angles were identified. The fracture propagation and stress evolution characteristics of rock with fractures at different angles were determined. The results reveal that compared to intact rocks, the peak stress, elastic modulus, peak strain, initiation stress, and damage stress of fractured rocks with different fracture angles around holes are lower. As the fracture angle increases, the gap in mechanical properties between the defective rock and the intact rock gradually decreased. In the force chain diagram, the compressive stress concentration range of the combined defect of cracks and holes starts to decrease, and the model is gradually destroyed as the tensile stress range gradually increases. When the peak stress is reached, the acoustic emission energy is highest and the rock undergoes brittle damage. Through a comparative study using laboratory tests, the results of laboratory real rocks and numerical simulation experiments were verified and the macroscopic failure characteristics of the real and simulated rocks were determined to be similar. This study can help us correctly understand the mechanical properties of rocks with defects and provide theoretical guidance for practical rock engineering.

Environmental fatigue correction factor model for domestic nuclear-grade low-alloy steel

  • Gao, Jun;Liu, Chang;Tan, Jibo;Zhang, Ziyu;Wu, Xinqiang;Han, En-Hou;Shen, Rui;Wang, Bingxi;Ke, Wei
    • Nuclear Engineering and Technology
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    • v.53 no.8
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    • pp.2600-2609
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    • 2021
  • Low cycle fatigue behaviors of SA508-3 low-alloy steel were investigated in room-temperature air, high-temperature air and in light water reactor (LWR) water environments. The fatigue mean curve and design curve for the low-alloy steel are developed based on the fatigue data in room-temperature and high-temperature air. The environmental fatigue model for low-alloy steel is developed by the environmental fatigue correction factor (Fen) methodology based on the fatigue data in LWR water environments with the consideration of effects of strain rate, temperature, and dissolved oxygen concentration on the fatigue life.

Improving the Viability of Freeze-dried Probiotics Using a Lysine-based Rehydration Mixture

  • Arellano, Karina;Park, Haryung;Kim, Bobae;Yeo, Subin;Jo, Hyunjoo;Kim, Jin-Hak;Ji, Yosep;Holzapfel, Wilhelm H.
    • Microbiology and Biotechnology Letters
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    • v.49 no.2
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    • pp.157-166
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    • 2021
  • The probiotic market is constantly continuing to grow, concomitantly with a widening in the range and diversity of probiotic products. Probiotics are defined as live microorganisms that provide a benefit to the host when consumed at a proper dose; the viability of a probiotic is therefore of crucial importance for its efficacy. Many products undergo lyophilization for maintaining their shelf-life. Unfortunately, this procedure may damage the integrity of the cells due to stress conditions during both the freezing and (vacuum-) drying process, thereby impacting their functionality. We propose a lysine-based mixture for rehydration of freeze-dried probiotics for improving their viability during in vitro simulated gastric and duodenum stress conditions. Measurement of the zeta potential served as an indicator of cell integrity and efficacy of this mixture, while functionality was estimated by adhesion to a human enterocyte-like Caco-2 cell-line. The freeze-dried bacteria exhibited a significantly different zeta potential compared to fresh cultures; however, this condition could be restored by rehydration with the lysine mixture. Recovery of the surface charge was found to influence adhesion ability to the Caco-2 cell-line. The optimum lysine concentration of the formulation, designated "Zeta-bio", was found to be 0.03 M for improving the viability of Lactiplantibacillus plantarum Lp-115 by up to 13.86% and a 7-strain mixture (400B) to 41.99% compared to the control rehydrated with distilled water. In addition, the lysine Zeta-bio formulation notably increased the adherence ability of lyophilized Lp-115 to the Caco-2 cell-line after subjected to the in vitro stress conditions of the simulated gastrointestinal tract passage.

Application of acyl-homoserine lactones for regulating biofilm characteristics on PAO1 and multi-strains in membrane bioreactor

  • Wonjung, Song;Chehyeun, Kim;Jiwon, Han;Jihoon, Lee;Zikang, Jiang;Jihyang, Kweon
    • Membrane and Water Treatment
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    • v.14 no.1
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    • pp.35-45
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    • 2023
  • Biofilms significantly affect the performance of wastewater treatment processes in which biodegradability of numerous microorganisms are actively involved, and various technologies have been applied to secure microbial biofilms. Understanding changes in biofilm characteristics by regulating expression of signaling molecules is important to control and regulate biofilms in membrane bioreactor, i.e., biofouling. This study investigated effects of addition of acyl-homoserine lactones (AHL) as a controllable factor for the microbial signaling system on biofilm formation of Pseudomonas aeruginosa PAO1 and multiple strains in membrane bioreactor. The addition of three AHL, i.e., C4-, C6-, and C8-HSL, at a concentration of 200 ㎍/L, enhanced the formation of the PAO1 biofilm and the degree of increases in the biofilm formation of PAO1 were 70.2%, 76.6%, and 72.9%, respectively. The improvement of biofilm formation of individual strains by C4-HSL was an average of 68%, and the microbial consortia increased by approximately 52.1% in the presence of 200 ㎍/L C4-HSL. CLSM images showed that more bacterial cells were present on the membrane surface after the AHL application. In the COMSTAT results, biomass and thickness were increased up to 2.2 times (PAO1) and 1.6 times (multi-strains) by C4-HSL. This study clearly showed that biofilm formation was increased by the application of AHL to individual strain groups, including PAO1 and microbial consortia, and significant increases were observed when 50 or 100 ㎍/L AHL was administered. This suggests that AHL application can improve the biofilm formation of microorganisms, which could yield an enhancement in efficiency of biofilm control, such as in various biofilm reactors including membrane bioreactor and bioflocculent systems in water/wastewater treatment processes.

Antibacterial Activity of JiYu-san Against Methicillin-resistant Staphylococcus aureus (Methicillin resistant staphylococcus aureus에 대한 지유산의 항균활성)

  • Hwang, Hae;Kang, Ok-Hwa;Kwon, Dong-Yeul
    • Korean Journal of Pharmacognosy
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    • v.53 no.2
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    • pp.87-95
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    • 2022
  • Methicillin resistance Staphylococcus aureus (MRSA) is a gram-positive bacterium, the most commonly isolated bacterial human pathogen. JiYu-san is one of the natural products used to treat diseases in the folk recipe. In this study, we investigated the antimicrobial activity of EtOH 70% extracts of JiYu-san (JYS) against MRSA. The antibacterial activity of JYS against MRSA strain was evaluated using minimum inhibitory concentration (MIC), checkerboard dilution test, and time-kill assay. The effect of JYS on the immune mechanism of MRSA was confirmed through cell membrane permeability tests and energy metabolism tests, and the antibacterial activity mechanism was performed using qRT-PCR and western blot. As a result, in the antibacterial test of JYS, the MIC was measured to be 1.9~1000 ㎍/mL, and synergistic or showed a partial synergistic effect. In addition, JYS showed antibacterial activity in a combination test with DCCD or TX-100. In a study on the mechanism of action of antibacterial activity, it was found that JYS suppressed MRSA resistance genes and proteins. These results suggest that JYS has antibacterial activity and provides great potential as a natural antibiotic by modulating the immune mechanism against MRSA.

A standardized method to study immune responses using porcine whole blood

  • Sameer-ul-Salam Mattoo;Ram Prasad Aganja;Seung-Chai Kim;Chang-Gi Jeong;Salik Nazki;Amina Khatun;Won-Il Kim;Sang-Myeong Lee
    • Journal of Veterinary Science
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    • v.24 no.1
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    • pp.11.1-11.14
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    • 2023
  • Background: Peripheral blood mononuclear cells (PBMCs) are commonly used to assess in vitro immune responses. However, PBMC isolation is a time-consuming procedure, introduces technical variability, and requires a relatively large volume of blood. By contrast, whole blood assay (WBA) is faster, cheaper, maintains more physiological conditions, and requires less sample volume, laboratory training, and equipment. Objectives: Herein, this study aimed to develop a porcine WBA for in vitro evaluation of immune responses. Methods: Heparinized whole blood (WB) was diluted (non-diluted, 1/2, 1/8, and 1/16) in RPMI-1640 media, followed by phorbol myristate acetate and ionomycin. After 24 h, cells were stained for interferon (IFN)-γ secreting T-cells followed by flow cytometry, and the supernatant was analyzed for tumor necrosis factor (TNF)-α. In addition, diluted WB was stimulated by lipopolysaccharide (LPS) and polyinosinic:polycytidylic acid (poly I:C), reference strain KCTC3557 (RS), field isolate (FI), of heat-killed (HK) Streptococcus suis, and porcine reproductive and respiratory syndrome virus (PRRSV). Results: The frequency of IFN-γ+CD3+ T-cells and concentration of TNF-α in the supernatant of WB increased with increasing dilution factor and were optimal at 1/8. WB TNF-α and interleukin (IL)-10 cytokine levels increased significantly following stimulation with LPS or poly I:C. Further, FI and RS induced IL-10 production in WB. Additionally, PRRSV strains increased the frequency of IFN-γ+ CD4-CD8+ cells, and IFN-γ was non-significantly induced in the supernatant of re-stimulated samples. Conclusions: We propose that the WBA is a rapid, reliable, and simple method to evaluate immune responses and WB should be diluted to trigger immune cells.

Characterization of Antibiotic Resistance of Aeromonas spp. and Pseudomonas spp. Isolated from Domestic Aquatic Animals (국내 수산생물로부터 분리된 Aeromonas spp. 및 Pseudomonas spp.의 항생제 내성에 관한 특성 분석)

  • Ye Ji Kim;Lyu Jin Jun;Young Juhn Lee;Ye Jin Ko;Yeong Eun Oh;Soo Ji Wo;Myoung Sug Kim;Joon Bum Jeong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.56 no.4
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    • pp.388-400
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    • 2023
  • Aeromonas spp. and Pseudomonas spp. are opportunistic pathogens widely distributed in the aquatic environment. To test the antibiotic susceptibility, the MIC of the 18 antibiotics mainly used in aquaculture were measured. Aeromonas spp. and Pseudomonas spp. straoms had different resistance patterns against most antibiotics. The MIC of tetracycline for four Aeromonas spp. strains (10.5%) was < 0.25 ㎍/mL. However, 0.5-4 ㎍/mL tetracycline inhibited most Pseudomonas spp. strains. The tet resistance performance of 14 genes including tet(B), tet(E), and tet(M) were investigated. Investigating, the tetracycline resistance gene of 38 Aeromonas spp. strains detected tet(A) in 21 strains (55.3%). Two Pseudomonas spp. strains showed high MIC values and no inhibition zone. tet gene analysis detected tet(D) in only one strain (5%).