• The Korean Journal of Pesticide Science
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• v.20 no.3
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• pp.181-188
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• 2016

BtPlus is a group of biopesticides that are made of Bacillus thuringiensis and immunosuppressant. A new BtPlus that exhibits high insecticidal activity against mosquito larvae has been investigated in control efficacy in field conditions and its environmental safety against aquatic system. This study assessed the control efficacy of BtPlus against mosquito larvae with two different application methods. In aerial spraying application (100 mL per $3.3m^2$), BtPlus was effective at 50% or above formulation concentrations to control mosquito larvae. For a direct application to aqueous mosquito habitat, a semi-field mimicking paddy rice field was constructed. In this condition, BtPlus showed 80% and 100% control efficacies at 0.1% and 0.2% concentrations, respectively. BtPlus also showed 40% mortality against adults at 0.1% concentration in 10% sugar bait. However, its control efficacies against adults were much less than against larvae. Safety assessment of BtPlus against ecosystem was evaluated using young carp (Cyprinus carpio), a water flea (Daphnia magna), and a honey bee (Apis mellifera). BtPlus did not give any adverse effects on these nontarget organisms. Based on these results, BtPlus can be applied to control mosquitoes by direct aqueous application to paddy rice field.

• Korean journal of applied entomology
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• v.54 no.2
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• pp.55-62
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• 2015

'Bt-Plus' has been developed by mixing spores of Bacillus thuringiensis (Bt) and culture broth of Xenorhabdus nematophila (Xn). Despite its high toxicity, it has some imitation to broaden its efficacy against diverse insect pest spectrum. This study focuses on enhancement of Bt-Plus toxicity against semi-susceptible insect, Spodoptera exitgua, by addition of Xn metabolites. Two main Xn metabolites, oxindole (OI) and benzylideneacetone (BZA), are known to enhance the Bt insecticidal activities. The addition of OI or BZA significantly increased Bt-Plus pathogenicity. However, when the freeze-dried Xn culture broth was added to Bt-Plus, much less amount was enough to enhance the toxicity compared to the amount of OI or BZA. An HPLC analysis indicated that there were more than 12 unidentifed bacterial metabolites in Xn culture broth. These suggest that there are potent biological response modifiers in Xn metabolites other than OI and BZA.

• Korean journal of applied entomology
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• v.52 no.2
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• pp.115-123
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• 2013

A microbial insecticide "Bt-Plus" has been developed to enhance an insecticidal efficacy of an entomopathogenic bacterium, Bacillus thuringiensis (Bt). However, its wettable powder formulation is not preferred by farmers and industry producers due to relatively high cost. This study aimed to develop a soluble concentrate formulation of Bt-Plus. To this end, an optimal mixture ratio of two bacterial culture broths was determined to be 5:4 (v/v) of Bt and Xenorhabdus nematophila (Xn) along with 10% ethanol preservative. In addition, Bt broth was concentrated by 10 times to apply the mixture at 1,000 times fold dilution. The resulting liquid formulation was sprayed on cabbage crop field infested by late instar larvae of the diamondback moth, Plutella xylostella. The field assay showed about 77% control efficacy at 7 days after treatment, which was comparable to those of current commercial biopesticides targeting P. xylostella. For storage test in both low and room temperatures, the liquid formation showed a relatively stable control efficacy at least for a month. To develop a quality control technique to exhibit a stable control efficacy of Bt-Plus, Bt spore density ($5{\times}10^{11}$ spores/mL) and eight active component concentrations of Xn bacterial metabolites in the formulation products have been proposed in this study.

• Korean journal of applied entomology
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• v.51 no.1
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• pp.47-58
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• 2012

Effect of a new crop protectant 'Bt-Plus' on natural enemies was analyzed in this study. Tested natural enemies included two parasitic species of $Aphidius$ $colemani$ and $Eretmocerus$ $eremicus$, and four predatory species of $Harmonia$ $axyridis$, $Orius$ $laevigatus$, $Amblyseius$ $swirskii$, and $Phytoseiulus$ $persimilis$. 'Bt-Plus' was formulated by combination of three entomopathogenic bacteria ($Xenorhabdus$ $nematophila$ (Xn), $Photorhabdus$ $temperata$ subsp. $temperata$ (Ptt), $Bacillus$ $thuringiensis$ (Bt)) and bacterial metabolite (BM). All three types of 'Bt-Plus' showed significantly higher toxicities against fourth instar $Plutella$ $xylostella$ larvae than Bt single treatment. Two types of bacterial mixtures ('Xn+Bt' and 'Ptt+Bt') showed little toxicity to all natural enemies in both contact and oral feeding assays. However, 'BM+Bt' showed significant toxicities especially to two predatory mites of $A.$ $swirskii$ and $P.$ $persimilis$. The acaricidal effects of different bacterial metabolites were evaluated against two spotted spider mite, $Tetranychus$ $urticae$. All six BM chemicals showed significant acaricidal effects. The BM mixture used to prepare 'Bt-Plus' showed a high acaricidal activity with a median lethal concentration at 218.7 ppm (95% confidence interval: 163.2 - 262.3). These toxic effects of bacterial metabolites were also proved by cytotoxicity test against Sf9 cells. Especially, benzylideneacetone, which was used as a main ingredient of 'BM+Bt', showed high cytotoxicity at its low micromolar concentration.

• Korean journal of applied entomology
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• v.62 no.4
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• pp.277-285
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• 2023

The onion moth, Acrolepiopsis sapporensis, was monitored in the farms cultivating the welsh onion, Allium fistulosum, using sex pheromone from transplantation to harvest. Two occurrence peaks were observed at early June and late July after the overwintering population. However, the population sizes were varied among different years and the cultivating environments. To effectively control A. sapporensis with microbial pesticides, different Bacillus thuringiensis strains were screened to select B. thuringiensis kurstaki (BtK). To enhance the insecticidal virulence of BtK, the culture broth of Photorhabdus temperata temperata (Ptt) was added to the BtK. This mixture of two entomopathogenic bacteria was called 'BtPlus', which was superior to BtK alone in the insecticidal virulence. The enhanced virulence was explained by the immunosuppressive activity of the secondary metabolites contained in the Ptt extract. The metabolites inhibited both cellular and humoral immune responses of A. sapporensis, resulting in the enhanced virulence of BtK. These results suggest that A. sapporensis occurs in the welsh onion fields and the resulting economic damage would be effectively prevented by BtPlus application.

• Korean journal of applied entomology
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• v.59 no.1
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• pp.41-54
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• 2020

Two mosquito species were collected in still-water near farming area in Andong, Korea. Based on morphological characters, these two mosquitoes were identified as Aedes koreicus and Culex vagans, respectively. DNA barcode analyses supported the identification. An entomopathogenic bacterium, Bacillus thuringiensis subsp. israelensis (BtI), exhibited insecticidal activities against the two mosquito species and its virulence was more potent than that of B. thuringiensis subsp. kurstaki. It has been known that the bacterial metabolites of Xenorhabdus spp. suppress insect immunity and enhance pathogenicity of B. thuringiensis. This study tested the effect of the bacterial culture broth of Xenorhabdus spp. on enhancing BtI pathogenicity. Among three Xenorhabdus spp., culture broth of X. ehlersii (Xe) was relatively effective to enhance BtI pathogenicity against both mosquito species. Indeed, organic extracts of Xe culture broth suppressed the hemocyte-spreading behavior, suggesting the presence of immunosuppressant in the culture broth. These results suggest a formulation of BtPlus by mixing BtI spore and Xe culture broth to be applied to control the two mosquito species.

• Korean journal of applied entomology
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• v.50 no.3
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• pp.171-178
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• 2011

Bacillus thuringiensis (Bt) is a bacterial biopesticide against insect pests, mainly lepidopterans. Spodoptera exigua and Plutella xylostella exhibit significant decreases in Bt susceptibility in late larval instars. To enhance Bt pathogenicity, we used a mixture treatment of Bt and other bacterial metabolites which possessed significant immunosuppressive activities. Mixtures of Bt with culture broths of Xenorhabdus nematophila (Xn) or Photorhabdus temperata ssp. temperata (Ptt) significantly enhanced the Bt pathogenicity against late larval instars. Different ratios of Bt to bacterial culture broth had significant pathogenicities against last instar P. xylostella and S. exigua. Five compounds identified from the bacterial culture broth also enhanced Bt pathogenicity. After determining the optimal ratios, the mixture was applied to cabbage infested by late instar P. xylostella or S. exigua in greenhouse conditions. A mixture of Bt and Xn culture broth killed 100% of both insect pests when it was sprayed twice, while Bt alone killed less than 80% or 60% of P. xylostella and S. exigua, respectively. Other Bt mixtures, including Ptt culture broth or bacterial metabolites, also significantly increased pathogenicity in the semi-field assays. These results demonstrated that the Bt mixtures collectively names "Bt-Plus" can be developed into potent biopesticides to increase the efficacy of Bt.