• Title/Summary/Keyword: Microbial Culture

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Characterization of Bacillus amyloliquefaciens DA12 Showing Potent Antifungal Activity against Mycotoxigenic Fusarium Species

  • Lee, Theresa;Park, Dami;Kim, Kihyun;Lim, Seong Mi;Yu, Nan Hee;Kim, Sosoo;Kim, Hwang-Yong;Jung, Kyu Seok;Jang, Ja Yeong;Park, Jong-Chul;Ham, Hyeonheui;Lee, Soohyung;Hong, Sung Kee;Kim, Jin-Cheol
    • The Plant Pathology Journal
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    • v.33 no.5
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    • pp.499-507
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    • 2017
  • In an attempt to develop a biological control agent against mycotoxigenic Fusarium species, we isolated Bacillus amyloliquefaciens strain DA12 from soil and explored its antimicrobial activities. DA12 was active against the growth of mycotoxigenic F. asiaticum, F. graminearum, F. proliferatum, and F. verticillioides both in vitro and in planta (maize). Further screening using dual culture extended the activity range of strain DA12 against other fungal pathogens including Botrytis cinerea, Colletotrichum coccodes, Endothia parasitica, Fusarium oxysporum, Raffaelea quercus-mongolicae, and Rhizoctonia solani. The butanol extract of the culture filtrate of B. amyloliquefaciens DA12 highly inhibited the germination of F. graminearum macroconidia with inhibition rate 83% at a concentration of $31.3{\mu}g/ml$ and 100% at a concentration of $250{\mu}g/ml$. The antifungal metabolite from the butanol extract was identified as iturin A by thin layer chromatography-bioautography. In addition, volatile organic compounds produced by DA12 were able to inhibit mycelial growth of various phytopathogenic fungi. The volatile compounds were identified as 2-heptanone, 5-methyl heptanone and 6-methyl heptanone by gas chromatography-mass spectrometry (GC-MS) analysis. These results indicate that the antagonistic activity of Bacillus amyloliquefaciens DA12 was attributable to iturin A and volatile heptanones, and the strain could be used as a biocontrol agent to reduce the development of Fusarium diseases and mycotoxin contamination of crops.

Detection of microbial organisms on Apis mellifera L. beehives in palm garden, Eastern Thailand

  • Sirikwan Dokuta;Sumed Yadoung;Peerapong Jeeno;Sayamon Hongjaisee;Phadungkiat Khamnoi;Khanchai Danmek;Jakkrawut Maitip;Bajaree Chuttong;Surat Hongsibsong
    • Journal of Ecology and Environment
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    • v.48 no.1
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    • pp.17-23
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    • 2024
  • Background: Honey bees play a crucial role in pollination and ecological balance. Apis mellifera L. colonies, especially those located in specific geographic regions, such as the palm garden in Eastern Thailand, are susceptible to potential threats from microbial contaminants. Understanding and detecting microbial organisms in these beehives is essential for the preservation of bee health, honey production, and the broader ecosystem. However, the problem of microbial infection and antibiotic-resistant bacteria is more severe and continuously increasing, resulting in a health, economic, and social crisis. The purpose of this study is to determine the prevalence of microorganisms in A. mellifera beehives in palm gardens in Rayong province, Eastern Thailand. Results: Ten swabs in transport media were swabbed and obtained from different parts of each beehive (1 swab per beehive), for a total of 10 hives. Traditional microbial culture-based methods, biochemical tests, and antimicrobial susceptibility (disc-diffusion) tests were used to detect microbial organisms and antibiotic resistance in bacteria. The swab tests from nine beehives resulted in the detection of Gram-positive bacteria (63.64%), Gram-negative bacteria (27.27%), and fungi/yeast (9.09%). These microorganisms are classified as a group of coagulase-negative Staphylococcus spp. and made up 40.91% of the bacteria discovered. Other bacteria found were Coryneform bacteria (13.64%), Pantoea spp. (13.64%), Bacillus spp. (9.09%), yeast (9.09%), glucose non-fermentative Gram-negative bacilli (9.09%), and Pseudomonas spp. (4.55%). However, due to the traditional culture-based and 0biochemical tests usually used to identify the microbial organisms in clinical specimens and the limitation of identifying some environmental microbial species, the results of the antimicrobial susceptibility test cannot reveal if the organism is resistant or susceptible to the drug. Nevertheless, drug-sensitive inhibition zones were formed with each antibiotic agent. Conclusions: Overall, the study supports prevention, healthcare, and public health systems. The contamination of microorganisms in the beehives may affect the quality of honey and other bee products or even the health of the beekeeper. To avoid this kind of contamination, it is therefore necessary to wear personal protective equipment while harvesting honey and other bee products.

Immunostimulation Effects of Cell Wall Components Isolated from Lactobacillus plantarum

  • TAE BOO CHOE;KANG, KWAN YUEB;SUNG HO PARK
    • Journal of Microbiology and Biotechnology
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    • v.4 no.3
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    • pp.195-199
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    • 1994
  • Immunostimulation effects of the cell wall components isolated from Lactobacillus plantarum were investigated by studying the macrophage s tumorcidal activity, splenocyte proliferation, anticomplementary activity and the inhibition of peritoneal tumor cell growth measured with ICR mice inoculated with sarcoma 180. The immunopotentiating cell wall components were a complex of peptidoglycan and exopolysaccharides. The tumorcidal activity of macrophage against Yacl and B16 tumor cells was enhanced when the cell wall components were added into the macrophage s culture medium. They also stimulated splenocytes to proliferate up to the same level as when the concanavalin A was added into the splenocyte's culture medium. The complementary activity was inhibited by 50% when the cell wall components were incubated with the sheep red blood cells treated with hemolysin and guinea pig complement. This result confirmed that the cell wall components had an antitumor effect, because the anticomplementary activity is usually accompanied by an antitumor activity at the same time. This fact was confirmed again by the inhibition of the growth of sarcoma 180 when the cell wall components were injected intraperitoneally into ICR mice inoculated with sarcoma 180. As a result, it is concluded that the cell wall components isolated from Lactobacillus plantarum had multifunctional immunostimulation effects in vitro and in vivo.

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Study of the Microbial and Chemical Properties of Goat Milk Kefir Produced by Inoculation with Taiwanese Kefir Grains

  • Chen, Ming-Ju;Liu, Je-Ruei;Lin, Chin-Wen;Yeh, Yu-Tzu
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.5
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    • pp.711-715
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    • 2005
  • One of the prerequisites for the successful implementation of industrial-scale goat kefir production is to understand the effects of different kefir grains and culture conditions on the microbial and chemical properties of the goat kefir. Thus, the objectives of the present study were to evaluate the characteristics of kefir grains in Taiwan on the microbial and chemical properties of goat milk kefir, as well as to understand the influence of culture conditions on production of medium chain-length triglycerides (MCT). Kefir grains were collected from households in northern Taiwan. Heat-treated goat milk was inoculated with 3-5% (V/W) kefir grains incubated at 15, 17.5, 20 or 22.5$^{\circ}C$ for 20 h, and the microflora count, ethanol content, and caproic (C6), caprylic (C8), and capric acid (C10) levels measured at 4 h intervals. Our results indicate that incubation with kefir grains results in 10$^6$-10$^7$ CFU/ml microflora count and 1.18 g/L of ethanol content at 20 h of fermentation. Incubation with 5% kefir grain at 20-22.5$^{\circ}C$ produces the highest MCT levels.

The Optimal Medium Composition for the Production of Microbial Cellulose by Acetobacter xylinum (Acetobacter xylinum에 의한 미생물 셀룰로오스의 생산을 위한 배지 최적조성)

  • Lee, Hei-Chan;Zhao, Xia
    • KSBB Journal
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    • v.11 no.5
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    • pp.550-556
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    • 1996
  • A complex medium was developed for the production of microbial cellulose by Acetobacter xylinum ATCC 23769. The optimum concentration of each nutrient for the production of microbial cellulose was determined to be 10g peptone, 20g yeast extract, 5g glucose, 1.56g Na2HPO4, 1.8g KH2PO4, 0.05g MgSO4, 0.002g FeCl3, 5g citric acid and 10 mL ethanol per liter. With synergistic effects of citric acid and ethanol, cellulose productivity achieved in developed medium was 0.446 gram of cellulose per gram glucose for static culture, which is much higher than reported values. Cell growth and the cellulose production in the developed medium under static culture was also investigated.

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Quality Stability of the Herb Pill Coated with Edible Oils Containing Rosemary Essential Oil (로즈마리를 첨가만 유지 코팅 생약제 환의 품질안정성)

  • Kwak, Yi-Sung;Choo, Jong-Jae
    • Journal of the Korean Society of Food Culture
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    • v.18 no.2
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    • pp.134-138
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    • 2003
  • Quality stability of the herb pill coated with edible oils containing rosemary was investigated. Herb pills were made of herb powders such as Panax ginseng, Cinnamomum cassia, Lycium chinense, Zyzyphus jujuba and Zingiber officinale. Rapeseed oil and lubriol were used as edible coating oil. After herb pills coated with edible oils with or without rosemary were stored at $40^{\circ}C$ for 180 days, the microbial viable cell counts and peroxide values(POV) of the herb pill were investigated. After 180 day storage, POVs of herb pills with only rapeseed oil or lubriol were 0.51 and 0.49 meq/kg, respectively. However, when rosemary was added in herb pills the POVs were decreased to 0.30 and 0.39 meq/kg, respectively. The addition of rosemary to the rapeseed oil and lubriol tended to decrease the microbial viable cell counts of the herb pill. The microbial viable cell counts of rapeseed oil and lubriol were 940 and 820CFU/g, respectively after 180 days of storage. However, these levels were suppressed to 720 and 640CFU/g by the resemary addition. On the other hand, the ginseng saponin content of herb pills was not affected by the rosemary addition during storage.

Quality Properties of Gangha-ju Liquor According to the Preparation Method (제조방법에 따른 강하주의 품질 특성)

  • Yu, Young-Ju;Jung, Soon-Teck
    • Journal of the Korean Society of Food Culture
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    • v.18 no.2
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    • pp.111-122
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    • 2003
  • This studies were performed to develop a Korean traditional folk liquor namely Gangha-ju has been prepared at Bosung district in Korea, and manufacturing conditions and anti-oxidation activity and anti-microbial activity of Gangha-ju were investigated. Ethyl-alcohol 20% and 30% Gangha-ju were brewed with glutinous rice wine, distilled liquor and 6 herbs of ginger, cinnamon, etc. Chemical and physical properties of 30% Gangha-ju were acidity 0.22, pH 4.31, amino acidity 3.26, transmittance 59 and conductivity $911\;{\mu}s/m$, and 20% Gangha-ju were 0.43, 4.20, 6.26, 62 and $924\;{\mu}s/m$. Volatile flavor compounds of ethyl alcohol, acetic acid, butanol, n-amyl alcohol, iso-pentyl alcohol, ethyl acetate, butyl acetate, acetaldehyde and furfural were detected, and main aroma compounds of Gangha-ju were isopentyl alcohol and ethyl acetate. Anti-oxidation activity by DPPH method was evaluated 31.32%, and nitrite scavenging effect was 31.79%. Anti-microbial activity against several microorganisms was pronounced strong activity over a wide range of test organisms, and Leuconostoc mesenteroids and Salmonella Ttyphimurium, Staphylococcus epidermidis were found to be more sensitive to Gangha-ju than Escherichia coli and Aspergillus flavus.

Shikimate Metabolic Pathway Engineering in Corynebacterium glutamicum

  • Park, Eunhwi;Kim, Hye-Jin;Seo, Seung-Yeul;Lee, Han-Na;Choi, Si-Sun;Lee, Sang Joung;Kim, Eung-Soo
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1305-1310
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    • 2021
  • Shikimate is a key high-demand metabolite for synthesizing valuable antiviral drugs, such as the anti-influenza drug, oseltamivir (Tamiflu). Microbial-based strategies for shikimate production have been developed to overcome the unstable and expensive supply of shikimate derived from traditional plant extraction processes. In this study, a microbial cell factory using Corynebacterium glutamicum was designed to overproduce shikimate in a fed-batch culture system. First, the shikimate kinase gene (aroK) responsible for converting shikimate to the next step was disrupted to facilitate the accumulation of shikimate. Several genes encoding the shikimate bypass route, such as dehydroshikimate dehydratase (QsuB), pyruvate kinase (Pyk1), and quinate/shikimate dehydrogenase (QsuD), were disrupted sequentially. An artificial operon containing several shikimate pathway genes, including aroE, aroB, aroF, and aroG were overexpressed to maximize the glucose uptake and intermediate flux. The rationally designed shikimate-overproducing C. glutamicum strain grown in an optimized medium produced approximately 37.3 g/l of shikimate in 7-L fed-batch fermentation. Overall, rational cell factory design and culture process optimization for the microbial-based production of shikimate will play a key role in complementing traditional plant-derived shikimate production processes.

Enhancing anaerobic digestion of vegetable waste and cellulose by bioaugmentation with rumen culture

  • Jo, Yeadam;Hwang, Kwanghyun;Lee, Changsoo
    • Membrane and Water Treatment
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    • v.10 no.3
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    • pp.213-221
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    • 2019
  • Anaerobic digestion (AD) has been widely used to valorize food waste (FW) because of its ability to convert organic carbon into $CH_4$ and $CO_2$. Korean FW has a high content of fruits and vegetables, and efficient hydrolysis of less biodegradable fibers is critical for its complete stabilization by AD. This study examined the digestates from different anaerobic digesters, namely Rs, Rr, and Rm, as the inocula for the AD of vegetable waste (VW) and cellulose (CL): Rs inoculated with anaerobic sludge from an AD plant, Rr inoculated with rumen fluid, and Rm inoculated with anaerobic sludge and augmented with rumen fluid. A total of six conditions ($3\;inocula{\times}2\;substrates$) were tested in serial subcultures. Biogas yield was higher in the runs inoculated with Rm than in the other runs for both VW (up to 1.10 L/g VS added) and CL (up to 1.05 L/g VS added), and so was biogas production rate. The inocula had different microbial community structures, and both substrate type and inoculum source had a significant effect on the formation and development of microbial community structures in the subcultures. The overall results suggest that the bioaugmentation with rumen microbial consortium has good potential to enhance the anaerobic biodegradability of VW, and thereby can help more efficiently digest high fiber-content Korean FW.

혐기성 PCE 탈염소화 미생물 농화 배양 및 미생물 군집 해석

  • 문부영;이태호;박태주
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.04a
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    • pp.332-336
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    • 2004
  • An anaerobic PCE(tetrachloroethylene) dechlorinating bacterial culture from a landfill soil was enriched and characterized. The enrichment culture could dechlorinate 60$\mu$mol/$m\ell$ of PCE during a month of incubation and cis-DCE(cis-dichloroethylene) was observed as a main product of PCE dechlorination. Microbial analysis of the dechlorinating enrichment culture by rising PCR-DGGE (Polymerase chain reaction-Denaturing gradient gel electrophoresis) method showed that at least three microorganisms were related to the anaerobic PCE dechlorination.

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