• Journal of Apiculture
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• v.32 no.1
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• pp.27-39
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• 2017

PCR-chip-based ultra-rapid multiplex PCRs for detection of six major infectious pathogens in honeybee were developed. The 6 kinds of major infectious pathogens in honeybee included Paenibacillus larvae causing American Foulbrood, Melissococcus plutonius causing European Foulbrood as bacteria, Ascosphaera apis (Chalkbrood), Aspergillus flavus (Stonebrood), Nosema apis and Nosema ceranae (Nosemosis) as fungi. The developed PCR-chip-based ultra-rapid multiplex PCR showed successful amplification for all six major pathogens in the presence of more than $10^3$ molecules. The time for confirming amplification (Threshold cycles; Ct-time) was about 7 minutes for two species, and about 9 minutes for four species. Total 40 cycles of PCR took 11 minutes 42 seconds and time for melting point analysis was 1 minute 15 seconds. Total time for whole PCR detection was estimated 12 minutes 57 seconds (40 cycles of PCR and melting point analysis). PCR-chip based ultra-rapid multiplex PCR using standard DNA substrates showed close to 100% accuracy and no false-amplification was found with honeybee genomic DNA. Ultra-rapid multiplex PCR is expected to be a fast and efficient pathogen detection method not only in the laboratory but also in the apiary field.

• Korean Journal of Food Science and Technology
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• v.42 no.3
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• pp.317-322
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• 2010

Aspergillus spp. is widely spread in the world on feeds and foods. They have been known to produce aflatoxins, which are mutagenic and carcinogenic to humans. The objective of this study was to determine the optimum gamma-irradiation dose for controling the growth of Aspergillus spp. to ensure safety of feeds. Four species fungal spore ($10^7$ spore/mL) exposed to 0, 1, 3, 5, 7 and 10 kGy of gamma radiation were inhibited in their growth and $AFB_1$ productivity by ${\geq}5\;kGy$. Meantime, the growth of fungal inoculated on feeds was inhibited at ${\geq}3\;kGy$. However, $AFB_1$ presented in aqueous solution was not be inactivated completely by ${\leq}10\;kGy$ irradiation. These results indicate that Aspergillus spp. on feeds could be controlled by 5 kGy gamma-irradiation but detoxification of $AFB_1$ demands a higher dose of gamma-irradiation (${\geq}10\;kGy$).

• Research in Plant Disease
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• v.17 no.3
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• pp.280-287
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• 2011

This investigation was undertaken to survey toxigenic fungal contamination of various rice samples in 93 rice processing complexes (RPC) in Korea. Rice was grown in 2009 and the samples were collected in 2010. Seven types of rice samples such as unhusked, brown, blue-tinged, discolored, polished, half-crushed, and rice husks were obtained from each RPC. One-hundred and five grains of each sample were placed on PDA plates after surface disinfection. The incidence of fungal contaminants was 26.8%. Aspergillus spp. was the most dominant fungal contaminants and Fusarium spp. was the most frequently occurred in samples. The heaviest Fusarium contamination was found in unhusked grain, rice husks, and bare blue-tinged rice and followed by colored rice whereas broken rice was the least contaminated. Regional difference of fungal contamination was distinctive. Fusarium incidence in the rice samples from southern region of Korea including Jeolla and Gyeongsang Provinces was higher than those from central region including Chungcheong, Gyeonggi, and Gangwon Provinces. In contrast to Fusarium spp., Aspergillus spp. and Penicillium spp. were dominated in brown and polished rice samples and their incidences were more severe in central region than southern region. The major contaminants shown more than 1% of kernels infected were Aspergillus (5.0%), Fusarium (2.0%), Alternaria (1.4%), Dreschlera (1.3%), Penicillium spp. (1.3%), and Nigrospora spp. (1.0%). Collectotrichum, Pyricularia, Myrothecium, Epicoccum, Cladosporium, Moniliella, Gloeocercospora, Chaeto- mium, Curvularia, Phialopora, Acremonium, Gliomastix, Trichoderma, Rhizopus, Phomopsis, Paecilomyces, Genicularia, Geotrichum, Acremoniella, Rhizoctonia, Phoma, Oidiodendran, and Candida spp. were among the rest observed at low incidence. The major contaminants of rice samples were well-known as toxigenic fungal genera so toxin producibility of these fungal isolates is necessary to be examined in future. It is also needed to study Myrothecium spp. on species level as it was detected for the first time in rice.

• Applied Biological Chemistry
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• v.43 no.2
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• pp.86-94
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• 2000

In order to develop the enzymatic hydrolysis system concerned with taste and flavor, strains having the high hydrolyzing activity on the soy protein were selected from some traditional Mejus. Two molds and one bacterium producing enzymes which were different in character of hydrolysis were isolated and identified. Leucine and azodye enzyme activities of both M4 and M5 were relatively high among in the isolated molds. And, leucine enzyme activity of B16 was the lowest in the isolated bacteria. These strains were isolated as microorganisms having a dissimilar hydrolysis pattern on the soy protein by enzymatic reactions. Mold M4 on the culture solid media was mycelium colors of white and its sclerotia colors were changed from white to black. According to the result of slide culture, radial conidial head, subclavate vesicle, conidia of subglobose, stipes of uncolored with smooth walls and metula and phialides were existed. Because M4 was taxonomically similar to the characteristics of Aspergillus oryzae (ahlburg) species, M4 was identified and named as Aspergillus oryzae M4.Mold M5 showed white and black mycelium on the MEA medium. Mold M5 colony exhibited grayish-green color and have long(7 mm) sporangiophores at slide culture. Sporangia became brownish-gray and the wall of larger sporangia was broken to form small collars, and smaller sporangia were fomed continually from large basal membrane. Columella is globose and hyaline, and sporangiospores are ellipsoidal of small diameter$(80\;{\mu}m)$. Because M5 was taxonomically similar to the Mucor circinelloides of zygomycetes, M5 was was identified and named as Mucor circinelloides M5. Bacteria B16 colony was opaque white, circular and lobate, and had rod shaped endospore. B16 was found positive in stain, catalase, ${\beta}-glucosidse$ and V-P tests. B16 was found to utilize D-fructose, ${\alpha}-D-glucose$, maltose, D-mannose, D-raffinose, stachyose and sucrose. By the morphological and physiological results, the characteristics of B16 was thought to correspond to that of Bacillus megaterium. However, fatty acid composition was similar to Paenibacillus marcerans, requiring further study for the definite identification. Accordingly, Bacteria B16 was provisionally classified and named as Bacillus megaterium B16.

• Journal of Food Hygiene and Safety
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• v.35 no.5
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• pp.438-446
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• 2020

Meju and nuruk (respectively soybean and malt) are traditional Korean fermentation starters that are vulnerable to contamination by harmful microorganisms such as aflatoxigenic fungi and their associated aflatoxins (AFs). In this study, Aspergillus spp. were isolated and identified from a total of 57 meju and 18 nuruk samples collected from Korean markets. Their potential aflatoxigenicity was investigated by examining the presence of three aflatoxin biosynthetic genes (aflO, aflP, and aflR) using multiplex polymerase chain reaction (mPCR) assays. Thereafter, aflatoxin production of isolates and the natural occurrence of AFs in meju and nuruk samples were analyzed by high-performance liquid chromatography (HPLC). A total of 177 Aspergillus isolates were identified and 130 isolates were obtained from meju samples. Of these, 25 isolates (19.2%) contained all three aflatoxin biosynthetic genes, and five (20%) of these isolates produced aflatoxins. Forty-seven of the Aspergillus isolates were obtained from nuruk samples, five of which (10.6%) expressed all three AF biosynthetic genes; however, none of these strains produced AFs. HPLC analysis showed that 88% (51/58) of the meju samples and 39% (7/18) of nuruk samples were not contaminated with AFs (below limit of detection). Among the isolates isolated from meju and nuruk, there were aflatoxigenic strains containing all three aflatoxin biosynthetic genes or producing aflatoxin in medium, but the frequency of aflatoxin contamination was low in the meju and nuruk samples.

• The Korean Journal of Mycology
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• v.15 no.3
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• pp.187-195
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• 1987

Two kinds of heat treatment method for the selective isolation of soil fungi to eliminate the commonest fungi and also to examine the vertical and seasonal distributions of the fungal population were applied to soil samples from two plots around Seoul area. The incubation method at $42^{\circ}C$ and heat treatment at $70^{\circ}C$ were used in this experiment. In the incubation method, the almost all the fungi isolated from two plots were mesophile, while the thermotolerant fungi was Aspergillus fumigatus and thermophilic fungi were Sporotrichum thermophile and Malbranchea pulchella var. sulfrea. The most dominant species isolated by this method was A. fumigatus. Nine genera and fourteen species were isolated from the two plots, and S. thermophile, Talaromyces ucrainicus,Malbranchea pulchella var. sulfrea were new to Korea. From the selection method by heat treatment at $70^{\circ}C$, ten genera and twenty species were isolated. Among these, the most fungi were also mesophile and thermotolerant fungus was A. fumigatus. The most dominant species isolated by this method was T. stipitatus, Talaromyces helicus var. major, Emericella nidulans var. nidulans, Chaetomium subspirale and Neosartorya fisheri var. fisheri were new to Korea. From the two isolation methods, it was found that the total number of soil fungi and frequency of species appeared including dominant ones were the highest at the soils of upper layer while the lowest at the soils of lower layer in its vertical distribution.

• Korean journal of food and cookery science
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• v.17 no.6
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• pp.565-570
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• 2001

Soibirhym(Portulace oleracea) was extracted by methanol and its antimicrobial activities against food spoilage or foodborne disease microorganisms were investigated by the paper disc method. The microorganisms used in this experiment included 5 species of bacteria(Escherichia coli, Pseudomonas aeruginosa, Salmonella typhimurium, Klebsiella Pneumoniae, Staphylococcus aureus) and 3 species of fungi(Fusarium solani, Aspergillus flavius, Penicillium citreonigrum). Soibirhym showed high antimicrobial activites against P. citreonigrum, P. aeruginosa and K. pneumoniae. Minimum inhibitory concentrations(MICs) on S. aureus, P. citreonigrum and K. pneumoniae were 200, 200 and 250 mg/㎖, respectively. In the methanol extracts from Soibirhym, 147 kinds of compound were separated by GC/MS. The extraction yields of phenolics, furans, alcohols, acids and esters, ketones, aldehydes, and miscellaneous compounds were 7.43%, 6.13%, 2.20%, 41.06%, 9.21%, 0.15% and 1.08%, respectively. Some antimicrobial compounds such as 2,3-dihydro-benzofuran, 4-hydroxy-3-methoxy-benzoic acid, 4-hydroxy benzeneethanol were detected in the methanol extract.

• The Korean Journal of Mycology
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• v.43 no.1
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• pp.13-19
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• 2015

Endophytic fungal strains were isolated from 5 aquatic plant species (Trapa japonica, Miscanthus sacchariflorus, Oenanthe javanica, Typha orientalis and Zizania latifolia) native to river wetland of tributary in Nakdong river. Total 34 strains were purely isolated, and then its internal transcribed spacer (ITS) regions were amplified. After that, phylogenetic analysis based on ITS sequences and deduction of diversity indices were done. Fungal isolates were belonged to 17 genera, concretely in Acremonium, Alternaria, Aspergillus, Cladosporium, Emericellopsis, Fusarium, Galactomyces, Leptosphaeria, Microsphaeropsis, Penicillium, Peyronellaea, Phoma, Pseudeurotium, Rhizomucor, Talaromyces, Trematosphaeria and Zalerion. Especially, fungal isolates were distributed intensively in genera Alternaria and Talaromyces. This study deals with the diversity of endophytic fungal species that showing symbiotic relationship with their host aquatic plants.

• Journal of Microbiology and Biotechnology
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• v.27 no.4
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• pp.678-684
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• 2017

The aim of this study was to elucidate the changes in the microbial community and biochemical properties of a traditional sweet paste during fermentation. PCR-denaturing gradient gel electrophoresis (DGGE) analysis showed that Aspergillus oryzae was the predominant species in the koji (the fungal mixture), and the majority of the fungi isolated belonged to two Zygosaccharomyces species in the mash. The bacterial DGGE profiles revealed the presence of Bacillus subtilis during fermentation, and Lactobacillus acidipiscis, Lactobacillus pubuzihii, Lactobacillus sp., Staphylococcus kloosi, and several uncultured bacteria were also detected in the mash after 14 days of main fermentation. Additionally, during main fermentation, amino-type nitrogen and total acid increased gradually to a maximum of $6.77{\pm}0.25g/kg$ and $19.10{\pm}0.58g/kg$ (30 days) respectively, and the concentration of reducing sugar increased to $337.41{\pm}3.99g/kg$ (7 days). The 180-day fermented sweet paste contained $261.46{\pm}19.49g/kg$ reducing sugar and its pH value remained at around 4.65. This study has used the PCR-DGGE technique to demonstrate the microbial community (including bacteria and fungi) in sweet paste and provides useful information (biochemical properties) about the assessment of the quality of sweet paste throughout fermentation.

• Mycobiology
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• v.42 no.3
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• pp.286-290
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• 2014

Fungi are the known sources of irritation associated with atopic diseases (e.g., asthma, allergic rhinoconjunctivitis, and atopic eczema). To quantitatively estimate their presence in the indoor environment of atopic dermatitis-inflicted child patient's houses (ADCPHs), the high-efficiency particulate air (HEPA) filters installed inside the air cleaners of three different ADCPHs were investigated for the presence of mold. The air cleaner HEPA filters obtained from the three different ADCPHs were coded as HEPA-A, -B, and -C, respectively, and tested for the presence of mold. The colony forming units (CFUs) corresponding to the HEPA-A, -B, and -C filters were estimated to be $6.51{\times}10^2{\pm}1.50{\times}10^2CFU/cm^2$, $8.72{\times}10^2{\pm}1.69{\times}10^2CFU/cm^2$, and $9.71{\times}10^2{\pm}1.35{\times}10^2CFU/cm^2$, respectively. Aspergillus, Penicillium, Alternaria, Cladosporium, Trichoderma, and other fungal groups were detected in the 2,494 isolates. The distribution of these fungal groups differed among the three filters. Cladosporium was the major fungal group in filters HEPA-A and -C, whereas Penicillium was the major fungal group in the filter HEPA-B. Nine fungal species, including some of the known allergenic species, were identified in these isolates. Cladosporium cladosporioides was the most common mold among all the three filters. This is the first report on the presence of fungi in the air cleaner HEPA filters from ADCPHs in Korea.