• Title/Summary/Keyword: B.thuringiensis

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Toxin Gene Analysis of Bacillus cereus and Bacillus thuringiensis Isolated from Cooked Rice (쌀밥에서 분리한 Bacillus cereus와 Bacillus thuringiensis의 독소유전자 분석)

  • Jeon, Jong-Hyuk;Park, Jong-Hyun
    • Korean Journal of Food Science and Technology
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    • v.42 no.3
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    • pp.361-367
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    • 2010
  • Bacterial contamination of cooked rice was analyzed to evaluate the microbial safety. Thirty raw rice samples were collected in Korea and cooked in an electric rice cooker. Mesophilic aerobe, food-poisoning Bacillus cereus group, and their toxin genes were determined on cooked rice. The percentage of total mesophilic aerobe based on 1-3 log CFU/g was 27% among the samples. Bacillus spp. in MYP selective medium was similar to the number of mesophilic aerobe, whileas Bacillus spp. was detected in most samples after enrichment. Thirty-seven isolates from 30 cooked rices were identified as B. thuringiensis, B. cereus, B. valismortis, B. pumilus, B. coagulans, B. licheniformis, Geobacillus stearothermophilus, and Brevibacillus laterosporus. Twenty isolates (54%), more than half of the isolates, were B. thuringiensis while nine (27%) were identified as B. cereus. All B. thuringiensis isolates possessed non-hemolytic toxin genes and interestingly, seven B. cereus among nine isolates possessed emetic toxin genes. More B. thuringiensis was present on the cooked rice than B. cereus and most B. cereus possessed emetic toxin genes rather than diarrheal toxin genes. Therefore, food-borne outbreak due to B.cereus on the cooked rice kept at room temperature might be examples of emetic food-poisoning.

Characterization of Two Type Crystal Proteins Produced by Transformed Bacillus thuringiensis NT0423 (형질전환된 Bacillus thuringiensis NT0423 균주에 의해 생산된 두가지 형태의 내독소 단백질의 특성)

  • 박현우;김호산;유용만;김상현;서숙재;강석권
    • Microbiology and Biotechnology Letters
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    • v.21 no.5
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    • pp.428-434
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    • 1993
  • Cloning and expression of two different crystal protein genes from transformed Bacillus thuringiensis were investigated. B. thuringiensis NT0423 is toxic to both Lepidopterous and Dipte-rous larvae. The pCG5 vector carrying crystal protein genes (mosquitocidal and hemolytic activity) of B. thurigiensis subsp. morrisoni PG-14 was transformed into B. thurigiensis NT0423. Transformant has expressed two type crystals of bipyramid from NT0423 and ovoid from pCG5 in one cell.

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Characterization of Bacillus thuringiensis subsp. tohokuensis CAB167 Isolate against Mosquito Larva (모기유충에 활성 있는 Bacillus thuringiensis subsp. tohokuensis CAB167 균주의 특성)

  • Kil, Mi-Ra;Kim, Da-A;Paek, Seung-Kyoung;Kim, Jin-Su;Choi, Su-Yeon;Jin, Da-Yong;Youn, Young-Nam;Hwang, In-Chon;Ohba, Michio;Yu, Yong-Man
    • Korean journal of applied entomology
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    • v.47 no.4
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    • pp.457-465
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    • 2008
  • Eight Bacillus thuringiensis strains activated against mosquito larva were compared their characterization. Spherical-shaped parasporal inclusion of B. thuringiensis subsp. tohokuensis CAB167 was observed by phase-contrast microscopy and scanning electron microscopy. $LC_{50}$ values of B. thuringiensis subsp. tohokuensis CAB167 against Culex pipiens molestus, Culex pipiens pallens, and Aedes aegyti were 173, 190 and 580 ng/ml, respectively. B. thuringiensis subsp. tohokuensis CAB167 had a parasporal inclusion containing 4 major protein components, for example, 135, 80, 49 and 28-kDa by SDS-PAGE. Otherwise, after trypsin digestion of parasporal inclusion, SDS-PAGE was showed new protease-resistant peptides at 72 and 63-kDa. Activated toxins of isolated CAB167 were different from other reference strains on a serological by immuno-diffusion test.

Selection and Antagonistic Mechanism of Bacillus thuringiensis BK4 against Fusarium Wilt Disease of Tomato (토마토시들음병의 생물학적 방제를 위한 토착길항세균 Bacillus thuringiensis BK4의 선발과 길항기작)

  • Jung Hee-Kyoung;Kim Jin-Rack;Kim Bo-Kum;Yu Tae-Shik;Kim Sang-Dal
    • Microbiology and Biotechnology Letters
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    • v.33 no.3
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    • pp.194-199
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    • 2005
  • In oder to select an antifungal substance-producing antagonistic bacterium against Fusarium oxysporum casuing fusarium wilt on tomato, strain BK4 was isolated from local soil of Gyeoungbuk and was identified as Bacillus thuringiensis by 16s rDNA analysis, biochemical test, and Mcirolog TM 3.0 System. The antibiotic of B. thuringiensis BK 4 was highly produced at $30^{\circ}C$ in nutrient broth (pH 9.0). The crude antibiotic was even stable at $121^{\circ}C$ and more stable at slight alkalic condition than acid condition. It was also remained $50{\%}$ activity at pH 3.0. B. thuringiensis BK4 showed the inhibition of spore germination and the biocontrol ability against F. oxysporum causing fusarium wilt of tomato in vivo test. According to these results, B. thuringiensis BK4 was enough to use with a microbial agent for biocontrol against fusarium wilt.

Characterization of bacillus thuringiensis isolate HL-15 from Korean soil (토양에서 분리한 bacillus thuringiensis의 특성)

  • 이형환;박미연;이창운
    • Korean Journal of Microbiology
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    • v.24 no.2
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    • pp.175-183
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    • 1986
  • The isolate HL-15 of Bacillus thuringiensis had common biochemical characteristics of the 23 serovarieties of B. thuringiensis. The isolates formed round endotoxin crystals which killed insect larvae, showed independent serological H antigen to the 23 serotypes, and contained two defferect DNA elements with over 100 Mdaltons of molecular weights.

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Various Enterotoxin and Other Virulence Factor Genes Widespread Among Bacillus cereus and Bacillus thuringiensis Strains

  • Kim, Min-Ju;Han, Jae-Kwang;Park, Jong-Su;Lee, Jin-Sung;Lee, Soon-Ho;Cho, Joon-Il;Kim, Keun-Sung
    • Journal of Microbiology and Biotechnology
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    • v.25 no.6
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    • pp.872-879
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    • 2015
  • Many strains of Bacillus cereus cause gastrointestinal diseases, and the closely related insect pathogen Bacillus thuringiensis has also been involved in outbreaks of diarrhea. The diarrheal diseases are attributed to enterotoxins. Sixteen reference strains of B. cereus and nine commercial and 12 reference strains of B. thuringiensis were screened by PCR for the presence of 10 enterotoxigenic genes (hblA, hblC, hblD, nheA, nheB, nheC, cytK, bceT, entFM, and entS), one emetogenic gene (ces), seven hemolytic genes (hlyA, hlyII, hlyIII, plcA, cerA, cerB, and cerO), and a pleiotropic transcriptional activator gene (plcR). These genes encode various enterotoxins and other virulence factors thought to play a role in infections of mammals. Amplicons were successfully generated from the strains of B. cereus and B. thuringiensis for each of these sequences, except the ces gene. Intriguingly, the majority of these B. cereus enterotoxin genes and other virulence factor genes appeared to be widespread among B. thuringiensis strains as well as B. cereus strains.

Resistance and Susceptibility of Diamondback Moth, Plutella xylostella Strains Collected from Different Region in Korea to Bacillus thuringiensis (국내 지역별 채집계통 및 감수성계통 배추좀나방에 대한 Bacillus thuringiensis 제품의 생물활성 비교)

  • Kim, Young-Rim;Cho, Min-Su;Oh, Se-Mun;Kim, Sung-Woo;Youn, Young-Nam;Yu, Yong-Man
    • The Korean Journal of Pesticide Science
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    • v.14 no.2
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    • pp.123-132
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    • 2010
  • Six populations of the diamondback moth, Plutella xylostella, were collected from the different national areas for resistance and reared in laboratory for two sensitive population. These populations of P. xylostella were examined the developed resistance against commercial products of Bacillus thuringiensis. There were 3 products with B. thuringiensis subsp. kurstaki including Tyuneup$^{(R)}$, Thuricide$^{(R)}$ and Geumulmang$^{(R)}$ and 2 products with B. thuringiensis subsp. aizawai including Tobagi$^{(R)}$ and Scorpion$^{(R)}$. The sensitive population of diamondback moths were provided from National Academy of Agricultural Science (NP) and Highland Agriculture Research Center (GR population) and field populations were caught from 6 different national areas. Resistance against Tyuneup$^{(R)}$ was developed 4.8 and 2.5 times in SP and HS compared with GR population of diamondback moth, respectively. In case of Geumulmang$^{(R)}$, it was developed 9.9 and 6.8 times in SP and NM population compared with NP population, respectively. Otherwise, Tobagi$^{(R)}$ was showed higher resistance in HS than any other population compared with GR population, however, Scorpion$^{(R)}$ that is a same strain with Tobagi$^{(R)}$, was showed only double resistance to SP population. It was supposed that the development of resistance to B. thuringiensis might be caused by the continuous application of the specific commercial product at the specific area. So, we need to use the commercial products of B. thuringiensis in rotation with different B. thuringiensis strains. In the other hand, when HS population with highest resistance were reared in laboratory, their resistance ratio was rapidly dropped to 1.1 times at second generation. We have to examined the resistance mechanism of the diamondback moth to B. thuringiensis strains.

Development of Bacillus thuringensis var. kurstaki NRD 6-Stu 1 by Site-Directed Mutagenesis (위치지정 변이기법을 이용한 Bacillus thuringiensis var. kurstaki NRD 6-Stu 1의 육성)

  • Lee, Jong-Su
    • The Journal of Natural Sciences
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    • v.6 no.1
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    • pp.41-48
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    • 1993
  • A cloned delta-endotoxin gene from Bacillus thuringiensis var. kurstaki NRD 6 was mutated at N-terminal toxic portion of the protoxin by site-directed mutagenesis. A Stu 1 restriction site was created in protoxin region of the mutant B.t.kurstaki NRD 6-Stu 1 which changed A to C in 177 region of the protoxin(silent mutation). Antigenic property of crystalline inclusion (delta-endotoxin) of the mutant was no difference to intact strain by the western blot analysis and its toxidity threshold value was also showed 0.015~0.030ng against Choristoneura femiferana-1 insect, similar with 0.01~0.024ng of parent strain, but stronger than that of B.t.kurstaki NRD 5 (500~1000ng) and B.t.kurstaki NRD 4(none toxic).

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Isolation, characterization, and evaluation of Bacillus thuringiensis isolated from cow milk

  • Kweon, Chang-Hee;Choi, Sang-Yoon;Kwon, Hyog-Young;Kim, Eun-Hye;Kang, Hyun-Mi;Moon, Jin-San;Jang, Geum-Chag;Lee, Hee-Soo;Kang, Seung-Won;Kim, Jong-Man;Pyo, Suhkneung;Rhee, Dong-Kwon
    • Korean Journal of Veterinary Research
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    • v.52 no.3
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    • pp.169-176
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    • 2012
  • Probiotics colonize the intestines and exert an antibacterial effect on pathogens. Therefore, probiotics could be used as a preventive agent against lethal infections. To isolate probiotic microorganisms, 116 bacterial strains were isolated from healthy cow's milk and were subjected to Gram-stain, morphology and biochemical analyses, Vitek analysis, and 16S rRNA analysis. One of the strains identified as Bacillus (B.) thuringiensis 87 was found to grow very well at pH 4.0~7.0 and to be resistant to high concentrations of bile salts (0.3~0.9% w/v). B. thuringiensis was susceptible to the antibiotics used in the treatment of bovine mastitis, yet it exhibited an antimicrobial effect against Staphylococcus (S.) aureus 305. Moreover, it protected mice from experimental lethal infections of E. coli O55, Salmonella typhimurium 01D, and S. aureus 305 through a significant induction of interferon-${\gamma}$, even at four-week post-administration of B. thuringiensis. Although oral administration of B. thuringiensis 87 did not provide significant protection against these lethal challenges, these results suggest that B. thuringiensis 87 could be a feasible candidate as a probiotic strain.

Examination of the Production of Extracellular $\alpha$-Amylase by Bacillus thuringiensis, 19 serotypes (Bacillus thuringiensis, 19 혈청형의 세포외 $\alpha$-Amylase 생산 검색)

  • 이건주;박동왕;이형환;이영주
    • Microbiology and Biotechnology Letters
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    • v.16 no.5
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    • pp.348-351
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    • 1988
  • The production of extracellular $\alpha$-amylase by Bacillus thuringiensis 19 serotypes was examined by the hydrolysis test of starch. Thirteen serotypes produced the amylase. B. thuringiensis serovar thuringiensis alesti, kurstaki, sotto, kenya, israelensis, morrisoni, entomocidus, tolworthi, thompsoni, toumanoffi, pakistani, and indiana produced tee enzyme. The amylase produced by B. thuringiensis serovar israelensis showed highest activity around pH 6.7 to 7.2 and 55$^{\circ}C$ to $65^{\circ}C$. The high production medium of the amylase was composed of 1% bactopeptone, 0.3% beef-extract, 0.3% yeast ex-tract, 0.5% NaCl, 0.3% $K_2$HPO$_4$, 0.1% KH$_2$PO$_4$, 0.2% Soluble Starch, 0.012% CaCl$_2$.2$H_2O$$_2$, 0.005% MnCl$_2$, and 0.03% MgCl$_2$.7$H_2O$. The highest production of the enzyme was observed at 4 hours culture in the soluble starch (0.6 units/$m\ell$) or glucose (0.43 units/$m\ell$) substrate.

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