• Title/Summary/Keyword: spore inactivation

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Evaluation of Disinfection Characteristics for Mixed Oxidants Produced by Electro-chemical Method (전기분해 방식에 의한 혼합산화제 소독 특성 평가)

  • Lee, Kyung-Hyuk;Lim, Jae-Lim;Lee, Doo-Jin;Kim, Seong-Su;Ahn, Hyo-Won
    • Journal of Korean Society of Water and Wastewater
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    • v.19 no.5
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    • pp.625-631
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    • 2005
  • Disinfection characteristics of mixed oxidants produced by mean of electro-chemical method were evaluated. Inactivation rate of B. subtilis spore on the mixed oxidants were similar with that of chlorine. Based on the experimental results, activation energy of mixed oxidants and chlorine with B. subtilis spore at pH 8.3 were calculated as 30.8, $34.7kJ{\cdot}mol^{-1}$ respectively. UV absorption spectrum of both chlorine and mixed oxidants present similar. Molar extinction coefficients of chlorine and mixed oxidants at 292nm, which is maximum absorption wavelength of $OCl^-$ were 357, $377M^{-1}cm^{-1}$, respectively. Disinfectant decay rate and formation rate of THM in the S WTP filtrate also shows insignificant difference for both mixed oxidants and chlorine. In consequent, it was confirmed that high portion of chlorine presents in the mixed oxidants.

Engineering CotA Laccase for Acidic pH Stability Using Bacillus subtilis Spore Display

  • Sheng, Silu;Jia, Han;Topiol, Sidney;Farinas, Edgardo T.
    • Journal of Microbiology and Biotechnology
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    • v.27 no.3
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    • pp.507-513
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    • 2017
  • Bacillus subtilis spores can be used for protein display to engineer protein properties. This method overcomes viability and protein-folding concerns associated with traditional protein display methods. Spores remain viable under extreme conditions and the genotype/phenotype connection remains intact. In addition, the natural sporulation process eliminates protein-folding concerns that are coupled to the target protein traveling through cell membranes. Furthermore, ATP-dependent chaperones are present to assist in protein folding. CotA was optimized as a whole-cell biocatalyst immobilized in an inert matrix of the spore. In general, proteins that are immobilized have advantages in biocatalysis. For example, the protein can be easily removed from the reaction and it is more stable. The aim is to improve the pH stability using spore display. The maximum activity of CotA is between pH 4 and 5 for the substrate ABTS (ABTS = diammonium 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate). However, the activity dramatically decreases at pH 4. The activity is not significantly altered at pH 5. A library of approximately 3,000 clones was screened. A E498G variant was identified to have a half-life of inactivation ($t_{1/2}$) at pH 4 that was 24.8 times greater compared with wt-CotA. In a previous investigation, a CotA library was screened for organic solvent resistance and a T480A mutant was found. Consequently, T480A/E498G-CotA was constructed and the $t_{1/2}$ was 62.1 times greater than wt-CotA. Finally, E498G-CotA and T480A/E498G-CotA yielded 3.7- and 5.3-fold more product than did wt-CotA after recycling the biocatalyst seven times over 42 h.

Some Environmental Factors Affecting Germination and Survival of Resting Spores of Plasmodioprora brassicae (배추무사마귀병균 휴면포자의 발아 및 생존에 미치는 몇가지 환경요인)

  • Kim, Choong-Hoe;Cho, Won-Dae;Kim, Hong-Mo
    • The Korean Journal of Pesticide Science
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    • v.4 no.4
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    • pp.66-71
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    • 2000
  • Effect of temperature on resting spore germination of Plasmodioprora brassicae was indirectly estimated based on examining temporal change of number of inactive resting spores. Resting spore germination was the highest at $28^{\circ}C$ reaching 55.6% and 82.5%, 24hr and 132hr after treatment, respectively. Optimum pH for resting spore germination was pH6, following pH7 and pH8, and the germination was inhibited at pH 4, and pH9. termination of resting spores was stimulated by root extracts of radish, Chinese cabbage and kidney bean, but inhibited by that of lettuce. Number of inactive resting spores was increased as temperature increases and time prolongs after temperature treatment. However, degree of inactivation of resting spores after 1hr at $40{\sim}65^{\circ}C$ was similar with $40{\sim}60%$, but rapidly increased to 91.5% at $70^{\circ}C$. When root galls were submerged in water, density of inactive resting spores was increased rapidly and reached 60.3% 9 days after treatment. Flooding of infested soil resulted in 30% reduction of survived resting spores 5 months later. Among the two registered fungicides, fluazinam was better for inactivation of resting spores than flusulfamide, but both fungicides were inferior to phosphoric acid.

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Studies on Inactivated Combined Vaccine of Bovine Anthrax and Blackleg (소의 탄저기종저 불활화 혼합백신에 관한 연구)

  • Jeon, Yun Seong
    • Korean Journal of Veterinary Research
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    • v.10 no.1
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    • pp.5-10
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    • 1970
  • Due to the fact that an inactivated anthrax vaccine may lark its immunogenicity and stability of immunogen a number of spore vaccines were exclusively used worldwide. In these studies a number of important factors were emphasized to achieve the following: selection of non or less allergic strain of anthrax bacillus, capsulation of bacteria. obtaining of non sporulating but vegetative organisms, adequate inactivation of B. anthraccis by means of formalin, adsorption of immunogen to aluminum hydroxide gel. Non or less allergic strains of anthrax bacillus which is inactivated with formalin was selected by a hyperimmunization and shock test in rabbits. Obtaining capsular material and vegetative immunogen, a virulent anthrax organisms were cultivated on sodium bicarbonate medium with of without adding of l-alanine in which B, anthracis grew luxuriantly without forming spores. Inactivation was carried out at $37^{\circ}C$ water bath for 3 days after the bacterial culture was mixed with formalin, in a final concentration of two per cent of formalin. Aluminum hydroxide gel was added to the mixture of anthrax and blackleg bacterin. Vaccines were injected guinea pig via subcutaneous or intramusoular route and challenged after three weeks and the possibilities of protection was tested. Throughout the studies. the above mentioned vaccines possibly protected the vaccinated guinea pigs more than 80 per cent compared to that of the controls. This experimental results strongly suggest that the vaccine may possibly applicable to bovine.

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Inactivation Mechanism of Bacillus subtilis Spores by Ethanol Extract of Torilis japonica Fruit

  • Cho, Won-Il;Cheigh, Chan-Ick;Choi, You-Jung;Jeong, Jeong-Yoon;Choi, Jun-Bong;Lee, Kang-Pyo;Cho, Seok-Cheol;Pyun, Yu-Ryang;Chung, Myong-Soo
    • Food Science and Biotechnology
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    • v.18 no.2
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    • pp.336-342
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    • 2009
  • To confirm the antimicobial mechanism of Torilis japonica, antimicrobial profile was observed on various spore conditions by combining 0.1% (3 mM) torilin with antimicrobial activity and 0.27% water fraction with germinants. A 75% ethanol extract of T. japonica fruit reduced Bacillus subtilis ATCC 6633 spore counts by 3 log cycles and reduced the vegetative cells to undetectable level (by about 6 log cycles) (both in terms of CFU/mL). Further fractionating the ethanol extract into n-hexane and water fractions revealed that the former reduced the spore count by 1 log cycle whereas the latter had no effect. The antimicrobial active compound was isolated and purified from the hexane layer, and identified as torilin ($C_{22}H_{32}O_5$). The water fraction of the ethanol layer did not show antimicrobial activity, whereas the antimicrobial effect of 0.1% (3 mM) torilin was significantly enhanced in the presence of the water fraction (0.27%). This result can be explained by synergistic effects of the water fraction containing considerable amounts of germinants such as L-alanine and K+ ions that triggered germination.

Elimination of Spore Bacteria in Beef by Gamma Irradiation (감마선 조사에 의한 우육오염 포자세균의 제거)

  • Kim, Sung;Yook, Hong-Sun;Choi, Cheong;Kim, Jeung-Ok;Byun, Myung-Woo
    • Journal of Food Hygiene and Safety
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    • v.13 no.3
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    • pp.294-298
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    • 1998
  • The effect of gamma irradiation on the survival of spore bacteria was investigated in frozen cells ($-18^{\circ}C$) with 0.1 M phosphate buffer and inoculated cells in beef. In the case of the frozen cells at log phase, the radiation $D_{10}\;and\;12D_{10}$ values were 0.29 kGy and 3.48 kGy in Bacillus subtilis, 0.39 kGy and 4.68 kGy in Bacillus cereus and 0.46 kGy and 5.52 kGy in Clostridium perfrigens. And inactivation factors were 6.52~10.34 and 10.87~17.24 at the dosage of 3 kGy and 5 kGy, respectively. The radiosensitivity of inoculated cells in beef showed the $D_{10}$ value of 0.59~0.76 kGy, the $12D_{10}$ value of 7.08~9.12 kGy, and inactivation factors of 3.95~8.47. The radiosensitivity of the frozen cells was higher than that of inoculated cells in beef.

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Photocatalytic disinfection of indoor suspended microorganisms (Escherichia coli and Bacillus subtilis spore) with ultraviolet light (광촉매와 UVA에 의한 실내 부유 미생물(E. coli 및 Bacillus. subtilis sp.) 살균 제거 연구)

  • Yoon, Young H.;Nam, Sook-Hyun;Joo, Jin-Chul;Ahn, Ho-Sang
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.15 no.2
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    • pp.1204-1210
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    • 2014
  • New control methods are proposed for indoor air quality by removing fine airborne dust-particles. As suspended fine dust-particles contain inorganic dust as well as fine organic bacteria, studies for simultaneous control of these contaminants are required. In this study, photocatalytic disinfection of indoor suspended microorganisms such as E. coli and Bacillus subtilis is performed by three types of photocatalysts with UVA irradiation. The UVA irradiation strength was controlled to the minimum $3{\mu}W/cm^2$, and ZnO, $TiO_2$, and ZnO/Laponite ball were used as the catalysts. The results indicate that E. coli was removed over 80 % after about 2 hours of reaction with UVA and all three types of photocatalysts, whereas only with UVA, around 50 % E. coli removal was obtained. Among the catalysts, ZnO/Laponite composite ball was found to have similar sterilizing capacity to $TiO_2$. However, in case of B. subtilis, which has thick cell wall in its spore state, disinfection was not effective under the low UVA irradiation condition, even with the catalysts. Further studies need to figure out the optimal UVA irradiation ranges as well as photocatalysts doses to control airborne dust, to provide healthy clean air environment.

Studies on Inactivated Combined Vaccine of Bovine Anthrax and Blackleg I. Preparation of Vaccine and Its Evaluation in Guinea Pigs (소의 탄저기종저 불활화 혼합백신에 관한 연구 I. 백신 제조와 기니픽에서의 효과시험)

  • Jeon, Yun Seong
    • Korean Journal of Veterinary Research
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    • v.12 no.1
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    • pp.71-75
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    • 1972
  • Due to the fact that an inactivated anthrax vaccine may show no or lower immunogenicity and stability, a number of spore vaccines were exclusively used worldwide. In these studies non or less allergic strain of anthrax bacillus was selected and made a capsulated vegetative organisms. Anthrax organisms of a virulent strain were cultivated on sodium bicarbonate medium with or without adding I-alanine in which B. anthracis grew luxuriantly without forming spores. Inactivation of the organisims was carried out at $37^{\circ}C$ water bath for 3 days after the bacterial culture was mixed with formalin in a final concentration of two per cent. Aluminum hydroxide gel was added to the mixture of anthrax and blackleg bacterin. Guinea pigs were injected with the vaccine via subcutaneous or intramuscular route and challenged after three weeks, and the possibilities of protection was tested. Throughout the studies, the vaccines possibly protected the vaccinated guinea pigs more than 80 per cent compared to that of the controls. This experimental results strongly suggest that the vaccine may possibly applicable to the prevention of bovine anthrax and blackleg.

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Effects of Ionizing Radiation on Postharvest Fungal Pathogens

  • Jeong, Rae-Dong;Shin, Eun-Jung;Chu, Eun-Hee;Park, Hae-Jun
    • The Plant Pathology Journal
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    • v.31 no.2
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    • pp.176-180
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    • 2015
  • Postharvest diseases cause losses in a wide variety of crops around the world. Irradiation, a useful nonchemical approach, has been used as an alternative treatment for fungicide to control plant fungal pathogens. For a preliminary study, ionizing radiations (gamma, X-ray, or e-beam irradiation) were evaluated for their antifungal activity against Botrytis cinerea, Penicillium expansum, and Rhizopus stolonifer through mycelial growth, spore germination, and morphological analysis under various conditions. Different fungi exhibited different radiosensitivity. The inhibition of fungal growth showed in a dose-dependent manner. Three fungal pathogens have greater sensitivity to the e-beam treatment compared to gamma or X-ray irradiations. The inactivation of individual fungal-viability to different irradiations can be considered between 3-4 kGy for B. cinerea and 1-2 kGy for P. expansum and R. stolonifer based on the radiosensitive and radio-resistant species, respectively. These preliminary data will provide critical information to control postharvest diseases through radiation.

A Distinguishing Staining Methods for Vitality of Kudoa (Kudoa septempunctata) spores in Muscle of Olive flounder (Paralichthys olivaceus) (넙치에 기생하는 쿠도아 점액포자충(Kudoa septempunctata) 의 활성 확인을 위한 염색법의 비교)

  • DO, Jeong-Wan;MOON, Sun Hwa;KIM, Min Ji;CHO, Mi-Young;JUNG, Sung Hee;LEE, Nam-Sil
    • Journal of Fisheries and Marine Sciences Education
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    • v.28 no.6
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    • pp.1822-1827
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    • 2016
  • Consumption of Paralichthys olivaceus in raw fish have been reported as the cause of outbreaks of food-born illness, and Kudoa septempunctata in muscle of Paralichtys olivaceus was suggested with the causative agent. For this reason, distinguish of vital and dead spore is important to study survivability of Kudoa septempunctata in human intestinal condition and in vitro inactivation of Kudoa septempunctata. In this reports, we suggest NR & MB (Neutral red and Methylene blue) staining method that is easier and simpler than the previously described HO & PI (Hoechst33342 and Propidium iodide) method according to a experimental condition.