• Journal of environmental and Sanitary engineering
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• v.13 no.2
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• pp.34-39
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• 1998

For more convenient and rapidly isolation of Bacillus thuringiensis subsp. israelensis(Bti), 1) heat treatment spore forming bacteria, 2) growth in enrichment culture media for Bacillus sp. and 3) selection of bacteria producing a lecithinase for Bacillus thuringiensis subsp. israelensis, were performed. Spore forming bacteria were counted 4.8 $\times $ 10$^{8}$cells/g soil on NAPGCY media, 9.2 $\times $ 10$^{7}$cells/g soil on NA media, and 3.6 $\times $ 10$^{8}$cells/g soil on NAAC media, respectively. Bacteria producing only a lecithinase were reached at 25.2% on medium contained egg york, bacteria only producing a delta-endotoxin were reached at 23.2% by phase contrast microscope, and bacteria producing a lecithinase & a delta-endotoxin simultaneously were reached at 13.7%. Bacillus thuringiensis which producing a lecithinase and a delta-endotoxin simultaneously among bacteria producing a lecithinase, were reached at 56.5%; A half of Bacillus thuringiensis was produced a delta-endotoxin, but not produced a lecithinase. Among 8 isolates of Bacillus thuringiensis, two strain of Bti which has a mosquito-cidal toxin, were detected by PCR using a specific primer of $\delta $-endotoxin gene from Bti.

• BMB Reports
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• v.34 no.2
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• pp.130-133
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• 2001

The amino acid sequences of 130 kDa and 72 kDa proteins responsible for the larvicidal activity of Bacillus thuringiensis var israelensis (Bti) were analyzed by hydrophobic moment plots. A search for highly amphiphilic $\alpha$-helices was made in these proteins using the helical hydrophobic moment as a criterion of amphiphilicity The protein segments of the largest hydrophobic moments were analyzed. In the present communication we report the surface seeking helices in 130 kDa and 72 kDa proteins of Bacillus thuringiensis var israelensis. It is assumed that the surface seeking segments may participate in one of the membrane-related functions of Bacillus thuringiensis.

• Korean Journal of Microbiology
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• v.23 no.2
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• pp.122-128
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• 1985

Bacillus thuringiensis serovar israelensis 4Q1 contains 8 different covalently closed-circular (CCC) plasmids of molecular weight 204, 267, 109, 103, 16, 7.6, 6.4, and 5.0kb. The three smallest plasmids, designated pBti6, and pBti8 may prove to be useful as cloning vectors because of thier size and ease of isolation. The three plasmids were incubated separately with 9 different restriction enzymes and 7 of the enzymes tested cleaved one or more of the plasmids. Plasmid pBti6 has a single site for Bg1 II, Pst I and Pvu II, two sites for Bc1 I and Eco RI, and five sites for Hind III. Plasmid pBti7 has a single site for Bam HI and Pst I, two sites for Hind III, and three sites for PvuII. Plasmic pBti8 has a single site for Bam HI, BelI and Hind III, two sites for Eco RI, and three sites for Bgl II and Pvu II. Composite restriction enzyme maps for pBti6, pBti7 and pBti8 were constructed. The sites of restriction enzyme cleavage were determined by single, double and partial digests of the plasmid DNA. All the restriction sites were aligned relative to the single Bgl II(pBti6), Pst I(pBti7) or Hind III(pBti8) site, respectively.

• Microbiology and Biotechnology Letters
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• v.13 no.3
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• pp.315-319
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• 1985

Delta-endotoxin crystals of B. thuringiensis var. kurstari and B. thuringiensis var. israelensis were purified by NaBr density gradient centrifigation and the wet weight of the BTK endotoxin was approximately 23.79％ of the cell wet weight and that of BTI was 25％. The shape of BTK crystal was bipyramidal, whose size was 1.7${\mu}{\textrm}{m}$ $\times$ 0.9${\mu}{\textrm}{m}$ and that of BTI was a spheroid, whose size was about 1.6$\times$0.45${\mu}{\textrm}{m}$. The molecular weight of BTK crystal protein was approximately 134,000 daltons and that of BTI was about 128,000 daltons.

• Microbiology and Biotechnology Letters
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• v.27 no.5
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• pp.359-363
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• 1999

To prevent appearance of resistant mosquitoes against $\delta$-endotoxin of bacillus thuringiensis subsp. israelensis (Bti) in field, a mosquitocidal Bacillus thuringiensis strain 21-2(Bt21-2) producing a new type of $\delta$-endotoxin was isolated. The strain Bt 21-2 belongs to H serotype 43, B. thuringiensis subsp. guiyangiensis (Btg). The $\delta$-endotoxins from the strain Bt 21-2 and the strain Bti were a cuboid shape morphologically, but the $\delta$-endotoxin of the strain Bt 21-2 was composed of 150, 90 and 70kDa proteins on SDS-PAGE, and the antigenicity of $\delta$-endotoxin of the strain Bt 21-2 was different from that of the strain Bti on immunoblot. The $\delta$-endotoxin gene of the strain Bt 21-2 was not amplified with specific primers of $\delta$-endotoxin gene (cry4A and cry4B) of the strain Bti on PCR.

• Microbiology and Biotechnology Letters
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• v.18 no.3
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• pp.301-304
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• 1990

In the mosquitocidal delta-endotoxins from Bacillus thuringiensis subsp, isruelensis and B. thuringiensis subsp. darmstudiensis 73E10-2, were contained an immunologically homologous protein. The homologous protein was confirmed from Ouchterlony test, irnmuno-electrophoresis, and enzyme linked immunoassay by polyclonal antibodies against the delta-endotoxins of both strains.

• The Korean Journal of Mycology
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• v.30 no.1
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• pp.50-55
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• 2002

Biological control against mushroom fly, Lycoriella mali, was performed by using Bacillus thuringiensis subsp. israelensis Bti-D and Bti-U, isolated from dead mushroom fly in oyster mushroom houses. Control values of the bacterial strains Bti-D and Bti-U against L. mali in bottle culture of oyster mushroom were 74.4% and 64.2%, respectively, and the value in small tray culture were 75.8% and 56.8%, respectively. In the experiment to develop the mass, cheap media for Bti-D and Bti-U isolates, the Biji broth (bean curd residue, called Biji in Korean language) was selected as a culture medium for an inexpensive and mass cultivation by the measurement of optical density of the two bacteria grown in the different media tested. Insecticidal effect of the formulation contained different ingredients that were prepared by using the Bti-D strain cultured in the Biji broth was tested in tray and bottle culture of oyster mushroom. The WCS formulation that contained corn starch as bio-gel (86.4%) was more effective to control the mushroom fly than living cells (69.1%) in bottle culture of oyster mushroom. Moreover, insecticidal effect of the WCS formulation was improved when water of pH 8 was used for dilution of the formulation. Effect of the WCS formulation using water of pH 8 and chemicals, Zuron (dimillin) W.P. on the control of mushroom fly and the productivity of oyster mushroom was investigated in tray culture of oyster mushroom. The Zuron W.P. was more effective to control the mushroom fly than the WCS formulation. However, compared with no treatment, the productivity of the mushroom treated with the WCS formulation was improved than that of the mushroom with Zuron W.P.

• Korean journal of applied entomology
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• v.49 no.2
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• pp.151-158
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• 2010

Among 18 Bacillus thuringiensis isolates with spherical parasporal inclusion from soils, B. thuringiensis subsp. israelensis CAB199 was selected. It was showing over 90% mortality against Aedes aegypti and Culex pipiens moletus. It was confirmed that this B. thuringiensis subsp. israelensis CAB199 isolate also had a insecticidal activity against Culex inatomii that was occurred in the marsh. Because most of mosquito larva were primarily situated or shifted from under- to surface water, we need to select long floating formulations on surface water for controlling mosquito larva. It was tested the pesticidal and control effects in the laboratory and wetland with two formulation types of B. thuringiensis subsp. israelensis, for example, wettable power (WP) and suspension concentrate type (SC). Laboratory test showed that SC formulation type was relatively faster and more effective against 3 tested mosquito species, C. pipiens, Aedes aegypti, and C. inatomii. Otherwise, the control efficacy of SC formulation type was more rapidly appeared against C. inatomii in the wetland.

• Microbiology and Biotechnology Letters
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• v.26 no.4
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• pp.369-372
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• 1998

Microbial insecticide formulations were prepared by liquid and semi-solid fermentations using Bacillus thuringiensis subsp. kurstaki, HL-106 (BTK-HL106), B. thuringiensis subsp. israelensis HL-63 (BTI-HL63) and B. sphaericus 1593 (BS-1593) strains. The liquid fermentation medium contained molasses 2%, dextrose 1.5%, peptone 2%, D-xylose 0.025%, CaCl$_2$ 0.1%, K$_2$HPO$_4$ 0.1%, KH$_2$PO$_4$ 0.1%, MgSO$_4$$.$7H$_2$O 0.03%, FeSO$_4$$.$7H$_2$O 0.002%, ZnSO$_4$$.$7H$_2$O 0.02%. The composition of the semi-solid fermentation medium was rice bran 45.2%, zeolite 31%, yeast powder 0.02%, corn powder 5%, dextrose 3%, lime 0.3%, NaCl 0.06%, CaCl$_2$ 0.02%, and H$_2$O 15.42%. Insecticide formulations produced in the liquid fermentation named BTK-HL106, BTI-HL63 and BS-1593 pesticides and those in the semi-solid fermentation were designated as BTK-HL106-1, BTI-HL63-1 and BS-1593-1 pesticides, respectively. The number of spore (endotoxin crystals) was 2.65${\times}$10$\^$9/ spores per $m\ell$ in the BTK-HL106 and 3.5${\times}$10$\^$10/ in the BTK-HL106-1 3.8${\times}$10$\^$9/ spores in the BTI-HL63 and 7.0${\times}$10$\^$10/ in the BTI-HL63-1, and 7.5${\times}$10$\^$9/ in the BS-1593 and 1.4${\times}$10$\^$10/ in the BS-1593-1. The spores in the BS-1593 formulation was produced two times more than the other formulations. The spores in the BTI-HL63-1 were contained twice than those in the BTK-HL106-1, and five times than those in the BS-1593-1. The results indicated that spore (endotoxin crystals) productions in the semi-solid fermentation increased about ten times than those in the liquid fermentations. $LC_{50}$s of the BTI-HL63 and BS-1593 were 4.5 $\mu\textrm{g}$, and those of the BTI-HL63-1 and BS-1593-1 were 1.5 $\mu\textrm{g}$. $LC_{50}$ of the BTK-HL106 was 1.5 mg and that of the BTK-HL106-1 was 0.9 mg. The $LC_{50}$s of the formulations in the semi-solid fermentations showed about two to three times higher than those in the liquid fermentations.

• The Korean Journal of Mycology
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• v.30 no.1
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• pp.44-49
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• 2002

The study was conducted to isolate and identify insecticidal bacteria for biological control of larvae of mushroom fly, Lycoriella mali, which is one of serious pests to oyster mushrooms during its cultivation period. Among eight bacteria isolated from the soil in the oyster mushroom beds and the dead body of L. mali, two bacteria, Bti-D and Bti-U showed more toxicity with mortality rate than other six-bacteria isolates. The two bacteria showed more toxicity in three instar of the period of development of the mushroom fly than in other instar. Symptoms of the larvae of L. mali infected by the two bacteria developed as follows: at the early infection, the front middle gut changed color to light brown, the middle gut to brown, whole body to black brown, and eventually, the fly died. For the identification of these isolates, cultural and biochemical characteristics by Bergey's manual and Biolog system, cell morphology by TEM, endospore and endotoxin by phase-contrast microscope, and test using 33H antisera were examined. According to the results, these two isolates, Bti-D and Bti-U were identified as Bacillus thuringiensis subsp. israelensis respectively.