[논문]Prevalence and Toxin Characteristics of Bacillus thuringiensis Isolated from Organic Vegetables

Kim, Jung-Beom; Choi, Ok-Kyung; Kwon, Sun-Mok; Cho, Seung-Hak; Park, Byung-Jae; Jin, Na Young; Yu, Yong Man; Oh, Deog-Hwan

doi:10.4014/jmb.1703.03063

Prevalence and Toxin Characteristics of Bacillus thuringiensis Isolated from Organic Vegetables 원문보기

Journal of microbiology and biotechnology, v.27 no.8, 2017년, pp.1449 - 1456

Kim, Jung-Beom (Department of Food Science and Technology, Sunchon National University) , Choi, Ok-Kyung (Division of Agricultural Inspection, Gyeonggi-do Research Institute of Health and Environment) , Kwon, Sun-Mok (Division of Agricultural Inspection, Gyeonggi-do Research Institute of Health and Environment) , Cho, Seung-Hak (Division of Bacterial Disease Research, Center for Infectious Disease Research, Korea National Institute of Health) , Park, Byung-Jae (Department of Food Science and Biotechnology, School of Bio-convergence Science and Technology, Kangwon National University) , Jin, Na Young (Department of Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) , Yu, Yong Man (Department of Applied Biology, College of Agriculture and Life Sciences, Chungnam National University) , Oh, Deog-Hwan (Department of Food Science and Biotechnology, School of Bio-convergence Science and Technology, Kangwon National University)

Abstract ▼ AI-Helper

The prevalence and toxin characteristics of Bacillus thuringiensis isolated from 39 organic vegetables were investigated. B. thuringiensis was detected in 30 out of the 39 organic vegetables (76.9%) with a mean value of 2.60 log CFU/g. Twenty-five out of the 30 B. thuringiensis isolates (83.3%) showed insecticidal toxicity against Spodoptera exigua. The hblCDA, nheABC, and entFM genes were found to be the major toxin genes, but the ces gene was not detected in any of the tested B. thuringiensis isolates. The hemolysin BL enterotoxin was detected in all 30 B. thuringiensis isolates (100%). The non-hemolytic enterotoxin complex was found in 27 out of 30 B. thuringiensis isolates (90.0%). The B. thuringiensis tested in this study had similar toxin gene characteristics to B. cereus, which possessed more than one toxin gene. B. thuringiensis could have the potential risk of foodborne illness based on the toxin genes and toxin-producing ability.

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문제 정의

Thus, this study was performed to determine the prevalence of B. cereus and B. thuringiensis in organic vegetables and to investigate the toxigenic potential based on the occurrence of toxin genes and production ability of enterotoxins as well as insecticidal activity.

제안 방법

A PCR assay was conducted to detect the cry gene for confirmation of B. thuringiensis using a JinTech B. cereus PCR kit (Jinsung-UniTech Co., Korea) according to the manufacturer’s instructions.
cereus[53]. The ability to produce two enterotoxins and the emetic toxin was assessed to estimate the potential risk of foodborne illness caused by B. cereus and B. thuringiensis strains, using the BCET-RPLA, BDE-VIA, and Singlepath Emetic Tox Mrk kits (Tables 4 and 5). The emetic toxin was not detected in any of the B.
To estimate the prevalence of B. cereus and B. thuringiensis in organic vegetables, their biochemical identification was conducted using the BAKARA plate and Vitek-II system (Table 1). B.
To test the insecticidal activity, each B. thuringiensis strain was inoculated into nutrient broth (BD BBL Difco, USA) for overnight culture and then spread on five nutrient agar (KisanBio Co., Korea) plates and incubated at 30℃ for 5 days. The occurrence of autolysis in the plates was examined with a phase contrast microscope (BX51; Olympus Co.

대상 데이터

Thirty-nine organic vegetables (26 leafy vegetables, 6 flowerhead brassicase, 4 fruiting vegetables, and 3 root and tuber vegetables) were purchased from retail markets in Gyeonggi-do, South Korea between May and July in 2014. Twenty-five grams of each sample was mixed with 225 ml of buffered peptone water (Oxoid Ltd, UK) in a sterile filter stomacher bag and homogenized by using a BagMixer stomacher (Interscience, France) for 2 min.

이론/모형

thuringiensis from each sample were streaked on TSA (Oxoid) and incubated at 30℃ for 24 h. The DNA templates were extracted using the boiling method for the PCR assay according to the previously described protocol[26]. PCR amplification with a 20 μl reaction mixture consisting of AccuPower PCR PreMix (Bioneer Co.

성능/효과

In conclusion, the toxin production characteristics of B. thuringiensis isolates tested in this study were similar to those of B. cereus. These results suggest that B.
thuringiensis isolates. The NHE complex was found in all isolates of B. cereus (100.0%) and all of B. thuringiensis isolates (100.0%) without insecticidal activity, but 22 out of 25 (88.0%) B. thuringiensis isolates with insecticidal activity produced NHE toxin.
thuringiensis strains, using the BCET-RPLA, BDE-VIA, and Singlepath Emetic Tox Mrk kits (Tables 4 and 5). The emetic toxin was not detected in any of the B. cereus and B. thuringiensis isolates, whereas the HBL enterotoxin was detected in all of five B. cereus (100.0%) and 30 (100.0%) B. thuringiensis isolates. The NHE complex was found in all isolates of B.
0%, respectively. The percentage of occurrence of these toxin genes in B. thuringiensis isolates with insecticidal activity was 100.0%, 100.0%, 100.0%, 100.0%, 100.0%, 100.0%, 100.0%, 36.0%, and 0.0%, respectively. These results indicated that whereas hblCDA, nheABC, and entFM were found to be the major toxin genes, the ces gene was not detected in any of the tested B.
0%). The percentage of occurrence of these toxin genes in B. thuringiensis isolates without insecticidal activity was 100.0%, 80.0%, 80.0%, 100.0%, 100.0%, 100.0%, 100.0%, 20.0%, and 0.0%, respectively. The percentage of occurrence of these toxin genes in B.
0%, respectively. These results indicated that whereas hblCDA, nheABC, and entFM were found to be the major toxin genes, the ces gene was not detected in any of the tested B. cereus and B. thuringiensis isolates.
thuringiensis isolates, insecticidal assays were performed with Lepidoptera larvae. Twenty-five out of 30 B. thuringiensis isolates (83.3%) tested in this study showed insecticidal toxicity against Lepidoptera larvae. The B.

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