• Title/Summary/Keyword: Microbial Culture

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A Study on the Development of a Microbial Insecticide -(With special emphasis on formulation)- (미생물(微生物) 살충제(殺蟲劑)의 개발(開發)에 관(關)한 연구(硏究) -(제제화(製劑化)를 중심(中心)으로)-)

  • Lee, Jae-Koo;Kim, Ki-Cheol;Kim, Do-Young
    • Applied Biological Chemistry
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    • v.22 no.2
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    • pp.123-134
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    • 1979
  • For the purpose of developing a microbial insecticide utilizing Bacillus thuringiensis Berliner, research was done and the following results were obtained. 1) As the freeze-dried matter of the cocoon-cooked water discarded from the filature contains much crude protein(51.825%) and a lot of inorganic salts, it can make a good nutrition source for the culture cf B. thuringiensis Berliner. 2) Based on the suspensibility, formula F-5 turned out to be the most suitable for insecticidal use. Its composition includes 0.2 g of the cell-spore-crystal mixture, 25 g of 200-mesh kaolin, 2.5 g of New Kalgen-NX-150, and 2.5 g of glycerine admixed with 8 ml of distilled water and granulated in 80-mesh size. 3) All the components of F-5, F-6 and F-7 are identical except that the amounts of cell-spore-crystal mixture of F-5, F-6, and F-7 are 0.2 g, 0.4 g, and 0.6 g, respectively. Accordingly, their physical properties are almost all the same. 4) Formulas F-5, F-6, and F-7 exhibited an excellent toxicity to Anomis mesogona Walker, Dendrolimus spectabilis Butler, and Margaronia perspectalis Walker at the concentration of 5%. 5) Formulas F-8 and F-9 which contain $NaHCO_3$ as one of their components showed a remarkably reduced toxicity to Anomis mesogona Walker and Dendrolimus spectabilis Butler than F-6 which does not contain $NaHCO_3$. 6) A maximum of $2.97{\times}10^9$ spores per ml was obtained by incubating B. thuringiensis in M-3 which has a pH of 7.05 and comprises 0.2% of ammonium sulphate and 0.8% of glucose dissolved in the cocoon-cooked water, with aeration for 96 hours. 7) Formula F-6 exhibited a somewhat reduced toxicity to Anomis mesogona Walker and Dendrolimus spectabilis Butler, when stored at room temperature for 70 days after formulation and it is desirable to keep it in a dark and cold place. 8) In held applications, formula F-6 showed a good activity in controlling Monema flavescens Walker. Margaronia perspectalis Walker, and Macrosiphum ibarae Matsumura.

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Rumen Fermentation was Changed by Feed Inoculation Method in In Vitro (사료 접종 방법에 의한 in vitro 반추위 발효 상성 변화)

  • Yoo, Dae-Kyum;Moon, Joon-Beom;Kim, Han-Been;Yang, Sung-Jae;Park, Joong-Kook;Lee, Se-Young;Seo, Ja-Kyeom
    • ANNALS OF ANIMAL RESOURCE SCIENCES
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    • v.30 no.3
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    • pp.111-120
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    • 2019
  • The objective of this study was to investigate the effect of different feed inoculation method on rumen fermentation in an in vitro. Three experimental treatments were used: control (CON, direct dispersion of feed (2 g) in rumen fluid), combinations of direct dispersion (1 g) and nylon bag (DNB, pore size: 50 ㎛, 1 g), and nylon bag (NB, 2 g). An in vitro fermentation experiment was carried out using strained rumen fluid for 48 h incubation time and timothy was used as a substrate. At the end of the incubation, in vitro dry matter digestibility (IVDMD), in vitro neutral detergent fiber digestibility (IVNDFD), pH, volatile fatty acids (VFA), ammonia nitrogen (NH3-N), and microbial community were evaluated and gas production was estimated at 3, 6, 12, 24, 48 h incubation periods. Gas production was higher in CON than DNB and NB at 6 and 12 h incubation time (p<0.01). There were no differences in final gas production, pH, NH3-N concentration, total VFA production, and VFA profiles among treatments. The IVDMD was lowest in CON (p<0.01) but the IVNDFD was not differed by feed distribution methods. There were no significant differences in general bacteria and fungi. Protozoa count was highest in NB treatment among treatments (p<0.01). The abundance of cellulolytic bacteria, Ruminococcus flavefaciens and Fibrobacter succinogenes, was highest in the CON among treatments (p<0.01).

A Study on Dental Caries Activity Assessment from Saliva of Students of Dentistry College (I) (치과대학 대학생 타액의 치아 우식활성도 평가(I))

  • Son, Seung Hwa;Kim, Dong Ae;Park, Young Min
    • Journal of dental hygiene science
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    • v.13 no.2
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    • pp.182-190
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    • 2013
  • Composition of oral microbial flora is suggested to play a role in the maintenance of oral health. Among them, Streptococcus mutans plays an essential role in the formation of dental plaque and it is being noticed as incipient infective bacteria of dental caries. The purpose of this study was to prepare the basic data about distribution of Streptococcus mutans detected from saliva by measuring colony density. It is well known that smoking as well as drinking is a factor of dental caries, however there are few investigations about distribution of Streptococcus mutans either smoking or drinking. The materials of this study were collected from students of dentistry college with general characteristics obtained by self-administered questionnaires. Students were asked to gently chew the paraffin to get saliva for 1 minute. Bacitracin disc in the culture medium was activated for 15 minutes, and then sample was incubated in the activated medium at $37^{\circ}C$ incubator for 48 hours. Streptococcus mutans colonization per 1 ml saliva was measured by the number of criteria. The severe levels of dental caries activity were significantly increased in the case of smoking (from 5.6% to 10%) or drinking (form 0% to 7%), respectively. Also, dental caries activity increased according to smoking and drinking periods longer. However, using the SPSS analytical process (Mann-Whitney's U), no significant differences were observed between the smoking and the non-smoking or the drinking and non-drinking, respectively. These results obtained from dental caries activity of saliva suggest that distribution of Streptococcus mutans following smoking or drinking can be used as a significant information for oral health.

Evaluation of adenosine triphosphate testing for on-farm cleanliness monitoring compared to microbiological testing in an empty pig farrowing unit

  • Yi, Seung-Won;Cho, Ara;Kim, Eunju;Oh, Sang-Ik;Roh, Jae Hee;Jung, Young-Hun;Choe, Changyong;Yoo, Jae Gyu;Do, Yoon Jung
    • Journal of Animal Science and Technology
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    • v.62 no.5
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    • pp.682-691
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    • 2020
  • Careful cleaning and disinfection of pigpens is essential to prevent disease spread and avoid the resultant economic losses. Hygiene in pigpens is generally evaluated by visual monitoring supplemented with bacteriological monitoring, which includes counting the total aerobic bacteria (TAB) and/or fecal indicator bacteria (FIB). However, these methods present drawbacks such as time and labor requirements. As adenosine triphosphate (ATP) is ubiquitous in all living organisms including microorganisms, this study aimed to directly compare the results of microbial assessment and ATP quantification, and to suggest possible detailed application methods of the ATP test for hygiene evaluation in pigpens of a farrowing unit. Before and after standard cleaning procedures, samples were collected from the floor corner, floor center, and feeding trough of four pigpens at different time points. No FIB were detected and both the TAB and ATP levels were significantly decreased in the floor center area after cleaning. FIB were continuously detected after cleaning and disinfection of the floor corners, and there was no significant ATP level reduction. The feeding trough did not show any significant difference in these values before and after cleaning, indicating insufficient cleaning of this area. The levels of TAB and ATP after cleaning were significantly correlated and the average ATP value was significantly lower in the absence of FIB than in their presence. In the absence of standard references, a more thorough hygiene management could be achieved evenly by supplementing cleaning or disinfection based on the lowest ATP results obtained at the cleanest test site, which in the present study was the floor center. Overall, these results indicate that the on-farm ATP test can be used to determine the cleanliness status, in addition to visual inspection, as an alternative to laboratory culture-based testing for the presence of microorganisms.

Quality Changes of Vegetables by different Cooking Methods (조리방법에 따른 채소류의 품질 변화)

  • Kim, Byeong-Cheol;Hwang, Jin-Young;Wu, Hyun-Jung;Lee, Se-Mi;Cho, Hyung-Yong;Yoo, Young-Mi;Shin, Hae-Hun;Cho, Eun-Kyung
    • Culinary science and hospitality research
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    • v.18 no.1
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    • pp.40-53
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    • 2012
  • The quality changes of vegetables including cucumbers, young pumpkins, carrots, radishes, onions by different cooking methods in $90^{\circ}C$ water, 0.5% NaCl solution, 1.0% NaCl solution and $400^{\circ}C$ superheated steam were investigated. The changes in pH of cooked vegetables were increased. Hunter color system values for L(lightness), a(redness) and b(yellowness) of the vegetables cooked in $400^{\circ}C$ supetbeated steam for 30 seconds were less compared with those in $90^{\circ}C$ water, 0.5% NaCl solution and 1.0% NaCl solution. The microbial growth was inhibited very effectively when the vegetables cooked in $400^{\circ}C$ superheated steam for 30 seconds. After cooked in $90^{\circ}C$ water, 0.5% NaCl solution, 1.0% NaCl solution, $400^{\circ}C$ superheated steam, vitamin C content decreased by different ratio depending on cooking methods and vegetable varieties. Based on the results of the quality changes of cooked vegetables, the superheated steam cooking method showed a positive effect on sterilization and maintaining pH, color, hardness of vegetable tissues, vitamin C content. The data obtained from this study could be utilized as basic information for the optimum conditions of superheated steam cooking to develope HMR(Home Meal Replacement).

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Biosurfactant as a microbial pesticide

  • Lee, Baek-Seok;Choi, Sung-Won;Choi, Ki-Hyun;Lee, Jae-Ho;Kim, Eun-Ki
    • 한국생물공학회:학술대회논문집
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    • 2003.04a
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    • pp.40-44
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    • 2003
  • Soil-borne infectious disease including Pythium aphanidermatum and Rhizoctonia solani causes severe damage to plants, such as cucumber. This soil-borne infectious disease was not controlled effectively by chemical pesticide. Since these diseases spread through the soil, chemical agents are usually ineffective. Instead, biological control, including antagonistic microbe can be used as a preferred control method. An efficient method was developed to select an antagonistic strain to be used as a biological control agent strain. In this new method, surface tension reduction potential of an isolate was included in the ‘decision factor’ in addition to the other factors, such as growth rate, and pathogen inhibition rate. Considering these 3 decision factors by a statistical method, an isolate from soil was selected and was identified as Bacillus sp. GB16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth was observed when Bacillus sp. GB16 was used. Therefore this strain was considered as plant growth promoting rhizobacteria (PGPR). The action of surface tension reducing component was deduced as the enhancement of wetting, spreading, and residing of antagonistic strain in the rhizosphere. This result showed that new selection method was significantly effective in selecting the best antagonistic strain for biological control of soil-borne infectious plant pathogen. The antifungal substances against P. aphanidermatum and R. solani were partially purified from the culture filtrates of Bacillus sp. GB16. In this study, lipopeptide possessing antifungal activity was isolated from Bacillus sp. GB16 cultures by various purification procedures and was identified as a surfactin-like lipopeptide based on the Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), high performance liquid chromatography mass spectroscopy (HPLC-MS), and quadrupole time-of-flight (Q-TOF) ESI-MS/MS data. The lipopeptide, named GB16-BS, completely inhibited the growth of Pythium aphanidermatum, Rhizoctonia solani, Penicillium sp., and Botrytis cineria at concentrations of 10 and 50 mg/L, respectively. A novel method to prevent the foaming and to provide oxygen was developed. During the production of surface active agent, such as lipopeptide (surfactin), large amount of foam was produced by aeration. This resulted in the carryover of cells to the outside of the fermentor, which leads to the significant loss of cells. Instead of using cell-toxic antifoaming agents, low amount of hydrogen peroxide was added. Catalase produced by cells converted hydrogen peroxide into oxygen and water. Also addition of corn oil as an oxygen vector as well as antifoaming agent was attempted. In addition, Ca-stearate, a metal soap, was added to enhance the antifoam activity of com oil. These methods could prevent the foaming significantly and maintained high dissolved oxygen in spite of lower aeration and agitation. Using these methods, high cell density, could be achieved with increased lipopeptide productivity. In conclusion to produce an effective biological control agent for soil-borne infectious disease, following strategies were attempted i) effective screening of antagonist by including surface tension as an important decision factor ii) identification of antifungal compound produced from the isolated strain iii) novel oxygenation by $H_2O_2-catalase$ with vegetable oil for antifungal lipopeptide production.

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Purtification of Parasporal Protein Crystals of Bacillus thuringiensis (Bacillus thuringiensis의 내독소 단백질의 분리1)

  • Kim, Yeong-Hun;Kim, Sang-Hyeon;Gang, Seok-Gwon
    • Journal of Sericultural and Entomological Science
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    • v.33 no.1
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    • pp.32-36
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    • 1991
  • The study has been carried out to acquire some basic informations about Bacillus thuringiensis for developing the microbial pesticide. Three strains of Bacillus thuringiensis var. Kurstaki, dendrolimus and aizawai, were used in the experiments as follows. Growth characteristics of each strain were examined and parasporal protein crystals were isolated from the mixtures of spores and protein crystals by the new method of centrifugation in two-layer cushion of Renograffin using a fixed angle rotor. The results are as follows. 1. No difference was shown in growth characterestics among three strains of B. thuringiensis. In growth curves, all strains reached to exponential phase by 2 hr and stationary phase by 7-8 hr after inoculation. 2. The pH of the culture media during exponential growth stage decreased about 1.4 of a pH unit at the beginning of sporulation, but recovered during the early stage of sporulation and then remained nearly constant during the later stage. 3. As 10$m\ell$ sample was applied to two-layer cushion of Renograffin and then centrifuged for 1hr at 27,000g a fixed angle rotor, the purity and recovery ratio was 99.9% and 5.8%, respectively. It has been shown that the new method for the isolation of parasporal protein crystals was more efficient than any from the estabilished methods.

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Analysis of Microbial Community Structure in Soil and Crop Root System I. Analysis of Bacterial Community Structure in the Soil and Root System of Red Pepper and Tomato (토양과 작물근계의 미생물군집 구조해석 I. 고추 및 토마토 재배지 토양과 근계의 세균군집 구조해석)

  • Kim, Jong-Shik;Kwon, Soon-Wo;Lee, Seon-Ju;Jung, Beung-Gan;Song, Jae-Kyeong;Go, Soong-Ju;Ryu, Jin-Chang
    • Korean Journal of Soil Science and Fertilizer
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    • v.32 no.3
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    • pp.319-325
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    • 1999
  • A culture-dependent survey of bacterial community in the soil-root system of red pepper and tomato was conducted by dilution plate count method. The bacterial community within soil was not different from that of rhizoplane. However, the populations of fluorescent, pseudomonads were higher in rhizoplanes than in soils and higher in healthy rhizoplanes than in Phytophthora disease-infested rhizoplanes. The bacterial community of the pepper cropped soil and rhizoplanes was very similar to that of the tomato-cropped soil and rhizoplanes. Among 285 identified bacterial colonies, most colonies were belong to two groups by fatty acid analyses: 52% of the 285 colonies were belong to low G + C gram positive bacteria group. Bacillus spp. and 33% were belong to high G + C gram positive bacteria group. In order to use beneficial microorganisms to agro-ecosystem, these data of field trials should be intensively accumulated.

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Characterization of PAH-Degrading Bacteria from Soils of Reed Rhizosphere in Sunchon Bay Using PAH Consortia (순천만 갈대근권 토양으로부터 얻은 PAH 분해세균의 특성 분석)

  • Kim Sung-Hyun;Kang Sung-Mi;Oh Kye-Heon;Kim Seung-Il;Yoon Byoung-Jun;Kahng Hyung-Yeel
    • Korean Journal of Microbiology
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    • v.41 no.3
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    • pp.208-215
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    • 2005
  • This study was accomplished in order to collect fundamental data on microbial roles in recycling process of reed rhizosphere. Sunchon bay, which is considered as one of the marsh and mud environments severely affected by human activities such agriculture and fisheries, was selected as a model place. In our initial efforts, two bacterial consortia were obtained by enrichment culture using PAH mixtures containing anthracene, naphthalene, phenanthrene and pyrene as the sources of carbon and energy, and four pure bacteria capable of rapid degradation of PAH were isolated from them. Four strains designated as SCB1, SCB2, SCB6, and SCB7 revealed by morphological, physiological and molecular analyses were identified as Burkholderia anthina, Alcaligenes sp., Achromobacter xylosoxidans., and Pseudomonas putida, respectively with over $99{\%}$ confidence. Notably, Burkholderia anthina SCB1 and Alcaligenes sp. SCB2 were found to utilize anthracene and pyrene more quickly than naphthalene and phenanthrene, whereas Achromobacter xylosoxidans SCB6 and Pseudomonas putida SCB7 exhibited similar growth and degradation patterns except for pyrene. These facts suggest that the rhizosphere microorganisms capable of PAH degradation might be used to clean up the contamination sites with polycyclic aromatic hydrocarbons.

Isolation and Characterization of Bacillus subtilis MP56 with Antimicrobial Activity against MDR (Multi Drug Resistant) Strains (다약제내성균에 대한 항균 활성을 가지는 Bacillus subtilis MP56 균주의 분리 및 특성분석)

  • Park, Sungyong;Yoo, Jincheol;Seong, Chinam;Cho, Seungsik
    • Korean Journal of Microbiology
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    • v.49 no.1
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    • pp.90-94
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    • 2013
  • A new Bacillus strain designated as MP56 producing antimicrobial substance has been isolated from the mud flat of Korea. The strain MP56 was found to exhibit broad spectrum of antimicrobial activity against Gram-positive pathogenic microorganisms and MDR (multi drug resistant) strains. The 16S rRNA sequence revealed that the MP56 was closely related to Bacillus subtilis with 99.93% homology. The optimal medium composition for production of antimicrobial substance in the B. subtilis MP56 were 1% mannitol, 1% oat meal, 0.01% $CaCl_2$. Antimicrobial activity of the culture broth against different pathogenic strains was assessed using the antimicrobial spectrum. The result suggests that Bacillus strain MP56 produces high quality antimicrobial substance that might be very useful to control varieties of pathogenic microbial growth.